Product References
Arabidopsis cryptochrome 1 controls photomorphogenesis through regulation of H2A.Z deposition.
The Plant cell
Mao Z,Wei X,Li L,Xu P,Zhang J,Wang W,Guo T,Kou S,Wang W,Miao L,Cao X,Zhao J,Yang G,Zhang S,Lian H,Yang HQ
Light is a key environmental cue that fundamentally regulates plant growth and development, which is mediated by the multiple photoreceptors including the blue light (BL) photoreceptor cryptochrome 1 (CRY1). The signaling mechanism of Arabidopsis thaliana CRY1 involves direct interactions with CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1)/SUPPRESSOR OF PHYA-105 1 and stabilization of COP1 substrate ELONGATED HYPOCOTYL 5 (HY5). H2A.Z is an evolutionarily conserved histone variant, which plays a critical
Mon Jul 19 00:00:00 UTC 2021
Blue light-dependent interactions of CRY1 with GID1 and DELLA proteins regulate gibberellin signaling and photomorphogenesis in Arabidopsis.
The Plant cell
Xu P,Chen H,Li T,Xu F,Mao Z,Cao X,Miao L,Du S,Hua J,Zhao J,Guo T,Kou S,Wang W,Yang HQ
Cryptochromes are blue light photoreceptors that mediate various light responses in plants and mammals. In Arabidopsis (Arabidopsis thaliana), cryptochrome 1 (CRY1) mediates blue light-induced photomorphogenesis, which is characterized by reduced hypocotyl elongation and enhanced anthocyanin production, whereas gibberellin (GA) signaling mediated by the GA receptor GA-INSENSITIVE DWARF1 (GID1) and DELLA proteins promotes hypocotyl elongation and inhibits anthocyanin accumulation. Whether CRY1 co
Fri Aug 13 00:00:00 UTC 2021
Phytochrome B interacts with SWC6 and ARP6 to regulate H2A.Z deposition and photomorphogensis in Arabidopsis.
Journal of integrative plant biology
Wei X,Wang W,Xu P,Wang W,Guo T,Kou S,Liu M,Niu Y,Yang HQ,Mao Z
Light serves as a crucial environmental cue which modulates plant growth and development, and which is controlled by multiple photoreceptors including the primary red light photoreceptor, phytochrome B (phyB). The signaling mechanism of phyB involves direct interactions with a group of basic helix-loop-helix (bHLH) transcription factors, PHYTOCHROME-INTERACTING FACTORS (PIFs), and the negative regulators of photomorphogenesis, COP1 and SPAs. H2A.Z is an evolutionarily conserved H2A variant which
Tue Jun 01 00:00:00 UTC 2021
Extracellular Histones Inhibit Fibrinolysis through Noncovalent and Covalent Interactions with Fibrin.
Thrombosis and haemostasis
Locke M,Longstaff C
Histones released into circulation as neutrophil extracellular traps are causally implicated in the pathogenesis of arterial, venous, and microvascular thrombosis by promoting coagulation and enhancing clot stability. Histones induce structural changes in fibrin rendering it stronger and resistant to fibrinolysis. The current study extends these observations by defining the antifibrinolytic mechanisms of histones in purified, plasma, and whole blood systems. Although histones stimulated plasmino
Thu Apr 01 00:00:00 UTC 2021
Transcriptional Regulation of Lipid Catabolism during Seedling Establishment.
Molecular plant
Cai G,Kim SC,Li J,Zhou Y,Wang X
Mon Jul 06 00:00:00 UTC 2020
Division of Labor between PCNA Loaders in DNA Replication and Sister Chromatid Cohesion Establishment.
Molecular cell
Liu HW,Bouchoux C,Panarotto M,Kakui Y,Patel H,Uhlmann F
Concomitant with DNA replication, the chromosomal cohesin complex establishes cohesion between newly replicated sister chromatids. Several replication-fork-associated "cohesion establishment factors," including the multifunctional Ctf18-RFC complex, aid this process in as yet unknown ways. Here, we show that Ctf18-RFC's role in sister chromatid cohesion correlates with PCNA loading but is separable from its role in the&nb
Thu May 21 00:00:00 UTC 2020
Nanostructured recombinant protein particles raise specific antibodies against the nodavirus NNV coat protein in sole.
Fish & shellfish immunology
Thwaite R,Berbel C,Aparicio M,Torrealba D,Pesarrodona M,Villaverde A,Borrego JJ,Manchado M,Roher N
Nervous necrosis virus (NNV) reassortant strains RGNNV/SJNNV have emerged as a potent threat to the Mediterranean marine aquaculture industry, causing viral encephalopathy and retinopathy (VER) in Senegalese sole (Solea senegalensis). In this study, a cheap and practical vaccine strategy using bacterial inclusion bodies made of the coat protein of a virulent reassortant strain of this betanodavirus was devised. The nanostructured recombinant protein nanoparticles, VNNV-CNP, were administered wit
Wed Apr 01 00:00:00 UTC 2020
Metallochaperones Are Needed for Mycobacterium tuberculosis and Escherichia coli Nicotinamidase-Pyrazinamidase Activity.
Journal of bacteriology
Sheen P,Monsalve A,Campos J,Huerta R,Antiparra R,Arteaga H,Duran P,Bueno C,Kirwan DE,Gilman RH,Zimic M
Thu Jan 02 00:00:00 UTC 2020
A broad-spectrum antiviral molecule, QL47, selectively inhibits eukaryotic translation.
The Journal of biological chemistry
de Wispelaere M,Carocci M,Burri DJ,Neidermyer WJ,Olson CM,Roggenbach I,Liang Y,Wang J,Whelan SPJ,Gray NS,Yang PL
Small-molecule inhibitors of translation are critical tools to study the molecular mechanisms of protein synthesis. In this study, we sought to characterize how QL47, a host-targeted, small-molecule antiviral agent, inhibits steady-state viral protein expression. We demonstrate that this small molecule broadly inhibits both viral and host protein synthesis and targets a translation step specific to eukaryotic cells. We show that QL47 inhibits protein neosynthesis initiated by both canonical cap-
Fri Feb 07 00:00:00 UTC 2020
An expanded library of orthogonal split inteins enables modular multi-peptide assemblies.
Nature communications
Pinto F,Thornton EL,Wang B
Mon Mar 23 00:00:00 UTC 2020
Viral DNA Binding to NLRC3, an Inhibitory Nucleic Acid Sensor, Unleashes STING, a Cyclic Dinucleotide Receptor that Activates Type I Interferon.
Immunity
Li X,Deng M,Petrucelli AS,Zhu C,Mo J,Zhang L,Tam JW,Ariel P,Zhao B,Zhang S,Ke H,Li P,Dokholyan NV,Duncan JA,Ting JP
Immune suppression is a crucial component of immunoregulation and a subgroup of nucleotide-binding domain (NBD) leucine-rich repeat (LRR)-containing proteins (NLRs) attenuate innate immunity. How this inhibitory function is controlled is unknown. A key question is whether microbial ligands can regulate this inhibition. NLRC3 is a negative regulator that attenuates type I interferon (IFN-I) response by sequestering and attenuating stimulator of interferon genes (STING) activation. Here we report
Tue Mar 19 00:00:00 UTC 2019
Generation and Selection of a Panel of Pan-Filovirus Single-Chain Antibodies using Cell-Free Ribosome Display.
The American journal of tropical medicine and hygiene
Kunamneni A,Clarke EC,Ye C,Bradfute SB,Durvasula R
Filoviruses which include ebolaviruses and marburgvirus can cause outbreaks of highly lethal hemorrhagic fever. This disease causes significant morbidity and mortality in humans and non-human primates with human fatality rates reaching 90% during some outbreaks. Currently there is lack of licensed vaccines or antivirals for these viruses. Since early symptoms of filovirus infection mimic more common diseases there is a strong unmet public health and biodefense need for broad-spectrum filovirus r
Mon Jul 01 00:00:00 UTC 2019
An engineered mutant of a host phospholipid synthesis gene inhibits viral replication without compromising host fitness.
The Journal of biological chemistry
He G,Zhang Z,Sathanantham P,Zhang X,Wu Z,Xie L,Wang X
Viral infections universally rely on numerous hijacked host factors to be successful. It is therefore possible to control viral infections by manipulating host factors that are critical for viral replication. Given that host genes may play essential roles in certain cellular processes any successful manipulations for virus control should cause no or mild effects on host fitness. We previously showed that a group of positive-strand RNA viruses enrich phosphatidylcholine (PC) at the sites of viral
Fri Sep 20 00:00:00 UTC 2019
Functional characterization of a subtilisin-like serine protease from Vibrio cholerae.
The Journal of biological chemistry
Howell M,Dumitrescu DG,Blankenship LR,Herkert D,Hatzios SK
The causative agent of the human diarrheal disease cholera exports numerous enzymes that facilitate its adaptation to both intestinal and aquatic niches. These secreted enzymes can mediate nutrient acquisition biofilm assembly and interactions with its host. We recently identified a secreted serine protease IvaP that is active in infected rabbits and human choleric stool. IvaP alters the activity of several host and pathogen enzymes in the gut and along with other secreted proteases decreases bi
Fri Jun 21 00:00:00 UTC 2019
Affinity Purification of NF1 Protein-Protein Interactors Identifies Keratins and Neurofibromin Itself as Binding Partners.
Genes
Carnes RM,Kesterson RA,Korf BR,Mobley JA,Wallis D
Neurofibromatosis Type 1 (NF1) is caused by pathogenic variants in the NF1 gene encoding neurofibromin. Definition of NF1 protein-protein interactions (PPIs) has been difficult and lacks replication, making it challenging to define binding partners that modulate its function. We created a novel tandem affinity purification (TAP) tag cloned in frame to the 3' end of the full-length murine Nf1 cDNA (mNf1). We show that this cDNA is functional and expresses neurofibromin, His-Tag, and can correct p
Wed Aug 28 00:00:00 UTC 2019
Bacterial outer membrane proteins assemble via asymmetric interactions with the BamA β-barrel.
Nature communications
Doyle MT,Bernstein HD
The integration of β-barrel proteins into the bacterial outer membrane (OM) is catalysed by the β-barrel assembly machinery (BAM). The central BAM subunit (BamA) itself contains a β-barrel domain that is essential for OM protein biogenesis but its mechanism of action is unknown. To elucidate its function here we develop a method to trap a native Escherichia coli β-barrel protein bound stably to BamA at a late stage of assembly in vivo. Using disulfide-bond crosslinking we fin
Fri Jul 26 00:00:00 UTC 2019
Sumoylation regulates the stability and nuclease activity of Saccharomyces cerevisiae Dna2.
Communications biology
Ranjha L,Levikova M,Altmannova V,Krejci L,Cejka P
Dna2 is an essential nuclease-helicase that acts in several distinct DNA metabolic pathways including DNA replication and recombination. To balance these functions and prevent unscheduled DNA degradation Dna2 activities must be regulated. Here we show that Dna2 function is controlled by sumoylation. We map the sumoylation sites to the N-terminal regulatory domain of Dna2 and show that in vitro sumoylation of recombinant Dna2 impairs its nuclease but not helicase activity. In cells the total lev
Mon Apr 20 00:00:00 UTC 2020
Metformin Promotes Antitumor Immunity via Endoplasmic-Reticulum-Associated Degradation of PD-L1.
Molecular cell
Cha JH,Yang WH,Xia W,Wei Y,Chan LC,Lim SO,Li CW,Kim T,Chang SS,Lee HH,Hsu JL,Wang HL,Kuo CW,Chang WC,Hadad S,Purdie CA,McCoy AM,Cai S,Tu Y,Litton JK,Mittendorf EA,Moulder SL,Symmans WF,Thompson AM,Piwnica-Worms H,Chen CH,Khoo KH,Hung MC
Metformin has been reported to possess antitumor activity and maintain high cytotoxic T lymphocyte (CTL) immune surveillance. However, the functions and detailed mechanisms of metformin's role in cancer immunity are not fully understood. Here, we show that metformin increases CTL activity by reducing the stability and membrane localization of programmed death ligand-1 (PD-L1). Furthermore, we discover that AMP-activated protein kinase (AMPK) activated by metformin directly phosphorylates S195 of
Thu Aug 16 00:00:00 UTC 2018
Substrate relay in an Hsp70-cochaperone cascade safeguards tail-anchored membrane protein targeting.
The EMBO journal
Cho H,Shan SO
Membrane proteins are aggregation-prone in aqueous environments and their biogenesis poses acute challenges to cellular protein homeostasis. How the chaperone network effectively protects integral membrane proteins during their post-translational targeting is not well understood. Here biochemical reconstitutions showed that the yeast cytosolic Hsp70 is responsible for capturing newly synthesized tail-anchored membrane proteins (TAs) in the soluble form. Moreover direct interaction of Hsp70 with
Wed Aug 15 00:00:00 UTC 2018
Systematic overexpression study to find target enzymes enhancing production of terpenes in Synechocystis PCC 6803, using isoprene as a model compound.
Metabolic engineering
Englund E,Shabestary K,Hudson EP,Lindberg P
Of the two natural metabolic pathways for making terpenoids biotechnological utilization of the mevalonate (MVA) pathway has enabled commercial production of valuable compounds while the more recently discovered but stoichiometrically more efficient methylerythritol phosphate (MEP) pathway is underdeveloped. We conducted a study on the overexpression of each enzyme in the MEP pathway in the unicellular cyanobacterium Synechocystis sp. PCC 6803 to identify potential targets for increasing flux to
Sat Sep 01 00:00:00 UTC 2018
Protein Nanoparticles Made of Recombinant Viral Antigens: A Promising Biomaterial for Oral Delivery of Fish Prophylactics.
Frontiers in immunology
Thwaite R,Ji J,Torrealba D,Coll J,Sabés M,Villaverde A,Roher N
In the search for an eminently practical strategy to develop immunostimulants and vaccines for farmed fish we have devised recombinant viral antigens presented as "nanopellets" (NPs). These are inclusion bodies of fish viral antigenic proteins produced in . Soluble recombinant proteins are too labile to endure the environment and maintain full functionality and therefore require encapsulation strategies. Yet when they are produced as nanostructures they can withstand the wide range of gastroint
Wed Nov 20 00:00:00 UTC 2019
Cadmium favors F-actin depolymerization in rat renal mesangial cells by site-specific, disulfide-based dimerization of the CAP1 protein.
Archives of toxicology
Liu Y,Xiao W,Shinde M,Field J,Templeton DM
Cadmium is a toxic metal that produces oxidative stress and has been shown to disrupt the actin cytoskeleton in rat renal mesangial cells (RMC). In a survey of proteins that might undergo Cd2+-dependent disulfide crosslinking, we identified the adenylyl cyclase-associated protein, CAP1, as undergoing a dimerization in response to Cd2+ (5-40 µM) that was sensitive to disulfide reducing agents, was reproduced by the disulfide crosslinking agent diamide, and was shown by site-directed mutagen
Thu Mar 01 00:00:00 UTC 2018
The fibronectin ED-A domain enhances recruitment of latent TGF-β-binding protein-1 to the fibroblast matrix.
Journal of cell science
Klingberg F,Chau G,Walraven M,Boo S,Koehler A,Chow ML,Olsen AL,Im M,Lodyga M,Wells RG,White ES,Hinz B
Dysregulated secretion and extracellular activation of TGF-β1 stimulates myofibroblasts to accumulate disordered and stiff extracellular matrix (ECM) leading to fibrosis. Fibronectin immobilizes latent TGF-β-binding protein-1 (LTBP-1) and thus stores TGF-β1 in the ECM. Because the ED-A fibronectin splice variant is prominently expressed during fibrosis and supports myofibroblast activation we investigated whether ED-A promotes LTBP-1-fibronectin interactions. Using stiffness-tunea
Thu Mar 01 00:00:00 UTC 2018
A Combination of Two Human Monoclonal Antibodies Prevents Zika Virus Escape Mutations in Non-human Primates.
Cell reports
Keeffe JR,Van Rompay KKA,Olsen PC,Wang Q,Gazumyan A,Azzopardi SA,Schaefer-Babajew D,Lee YE,Stuart JB,Singapuri A,Watanabe J,Usachenko J,Ardeshir A,Saeed M,Agudelo M,Eisenreich T,Bournazos S,Oliveira TY,Rice CM,Coffey LL,MacDonald MR,Bjorkman PJ,Nussenzweig MC,Robbiani DF
Tue Nov 06 00:00:00 UTC 2018
Site-specific O-glycosylation of members of the low-density lipoprotein receptor superfamily enhances ligand interactions.
The Journal of biological chemistry
Wang S,Mao Y,Narimatsu Y,Ye Z,Tian W,Goth CK,Lira-Navarrete E,Pedersen NB,Benito-Vicente A,Martin C,Uribe KB,Hurtado-Guerrero R,Christoffersen C,Seidah NG,Nielsen R,Christensen EI,Hansen L,Bennett EP,Vakhrushev SY,Schjoldager KT,Clausen H
The low-density lipoprotein receptor (LDLR) and related receptors are important for the transport of diverse biomolecules across cell membranes and barriers. Their functions are especially relevant for cholesterol homeostasis and diseases including neurodegenerative and kidney disorders. Members of the LDLR-related protein family share LDLR class A (LA) repeats providing binding properties for lipoproteins and other biomolecules. We previously demonstrated that short linker regions between these
Fri May 11 00:00:00 UTC 2018
Structural insights into the architecture and membrane interactions of the conserved COMMD proteins.
eLife
Healy MD,Hospenthal MK,Hall RJ,Chandra M,Chilton M,Tillu V,Chen KE,Celligoi DJ,McDonald FJ,Cullen PJ,Lott JS,Collins BM,Ghai R
The COMMD proteins are a conserved family of proteins with central roles in intracellular membrane trafficking and transcription. They form oligomeric complexes with each other and act as components of a larger assembly called the CCC complex, which is localized to endosomal compartments and mediates the transport of several transmembrane cargos. How these complexes are formed however is completely unknown. Here, we have systematically characterised the interactions between human COMMD proteins,
Wed Aug 01 00:00:00 UTC 2018
Recombinant TSR1 of ADAMTS5 Suppresses Melanoma Growth in Mice via an Anti-angiogenic Mechanism.
Cancers
Renganathan B,Durairaj V,Kirman DC,Esubonteng PKA,Ang SK,Ge R
Inhibiting tumor angiogenesis is a well-established approach for anticancer therapeutic development. A Disintegrin-like and Metalloproteinase with ThromboSpondin Motifs 5 (ADAMTS5) is a secreted matrix metalloproteinase in the ADAMTS family that also functions as an anti-angiogenic/anti-tumorigenic molecule. Its anti-angiogenic/anti-tumorigenic function is independent from its proteinase activity but requires its first thrombospondin type 1 repeat (TSR1). However it is not known if recombinant T
Mon Jun 11 00:00:00 UTC 2018
Acylated-acyl carrier protein stabilizes the Pseudomonas aeruginosa WaaP lipopolysaccharide heptose kinase.
Scientific reports
Kreamer NNK,Chopra R,Caughlan RE,Fabbro D,Fang E,Gee P,Hunt I,Li M,Leon BC,Muller L,Vash B,Woods AL,Stams T,Dean CR,Uehara T
Phosphorylation of Pseudomonas aeruginosa lipopolysaccharide (LPS) is important for maintaining outer membrane integrity and intrinsic antibiotic resistance. We solved the crystal structure of the LPS heptose kinase WaaP, which is essential for growth of P. aeruginosa. WaaP was structurally similar to eukaryotic protein kinases and, intriguingly, was complexed with acylated-acyl carrier protein (acyl-ACP). WaaP produced by in vitro transcription-translation was insoluble unless acyl-ACP was pres
Thu Sep 20 00:00:00 UTC 2018
A Bacterial Type III Effector Targets the Master Regulator of Salicylic Acid Signaling, NPR1, to Subvert Plant Immunity.
Cell host & microbe
Chen H,Chen J,Li M,Chang M,Xu K,Shang Z,Zhao Y,Palmer I,Zhang Y,McGill J,Alfano JR,Nishimura MT,Liu F,Fu ZQ
Most plant bacterial pathogens rely on type III effectors to cause diseases. Although it is well known that the plant hormone salicylic acid (SA) plays an essential role in defense, whether the master regulator of SA signaling, NPR1, is targeted by any plant pathogen effectors is unknown. SA facilitates the reduction of cytosolic NPR1 oligomers into monomers, which enter the nucleus and function as transcriptional coactivators of plant defense genes. We show that SA promotes the interaction betw
Wed Dec 13 00:00:00 UTC 2017
Collective Force Regulation in Anti-parallel Microtubule Gliding by Dimeric Kif15 Kinesin Motors.
Current biology : CB
Reinemann DN,Sturgill EG,Das DK,Degen MS,Vörös Z,Hwang W,Ohi R,Lang MJ
During cell division the mitotic kinesin-5 Eg5 generates most of the force required to separate centrosomes during spindle assembly. However Kif15 another mitotic kinesin can replace Eg5 function permitting mammalian cells to acquire resistance to Eg5 poisons. Unlike Eg5 the mechanism by which Kif15 generates centrosome separation forces is unknown. Here we investigated the mechanical properties and force generation capacity of Kif15 at the single-molecule level using optical tweezers. We found
Mon Sep 25 00:00:00 UTC 2017
Fbxl4 Serves as a Clock Output Molecule that Regulates Sleep through Promotion of Rhythmic Degradation of the GABAA Receptor.
Current biology : CB
Li Q,Li Y,Wang X,Qi J,Jin X,Tong H,Zhou Z,Zhang ZC,Han J
The timing of sleep is tightly governed by the circadian clock which contains a negative transcriptional feedback loop and synchronizes the physiology and behavior of most animals to daily environmental oscillations. However how the circadian clock determines the timing of sleep is largely unclear. In vertebrates and invertebrates the status of sleep and wakefulness is modulated by the electrical activity of pacemaker neurons that are circadian regulated and suppressed by inhibitory GABAergic in
Mon Dec 04 00:00:00 UTC 2017
Therapeutic Effects of FGF23 c-tail Fc in a Murine Preclinical Model of X-Linked Hypophosphatemia Via the Selective Modulation of Phosphate Reabsorption.
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
Johnson K,Levine K,Sergi J,Chamoun J,Roach R,Vekich J,Favis M,Horn M,Cao X,Miller B,Snyder W,Aivazian D,Reagan W,Berryman E,Colangelo J,Markiewicz V,Bagi CM,Brown TP,Coyle A,Mohammadi M,Magram J
Fibroblast growth factor 23 (FGF23) is the causative factor of X-linked hypophosphatemia (XLH), a genetic disorder effecting 1:20,000 that is characterized by excessive phosphate excretion, elevated FGF23 levels and a rickets/osteomalacia phenotype. FGF23 inhibits phosphate reabsorption and suppresses 1α,25-dihydroxyvitamin D (1,25D) biosynthesis, analytes that differentially contribute to bone integrity and deleterious soft-tissue mineralization. As inhibition of ligand broadly modulates
Sun Oct 01 00:00:00 UTC 2017
Expression, Purification, and Properties of a Human Arachidonoyl-Specific Isoform of Diacylglycerol Kinase.
Biochemistry
Jennings W,Doshi S,Hota PK,Prodeus A,Black S,Epand RM
Diacylglycerol kinase ε (DGKε) catalyzes the phosphorylation of diacylglycerol producing phosphatidic acid. DGKε demonstrates exquisite specificity for the acyl chains of diacylglycerol. This contributes to the enrichment of particular acyl chains within the lipids of the phosphatidylinositol cycle. Phosphatidylinositol is highly enriched with 1-stearoyl-2-arachidonoyl which is important for maintaining cellular health. Dysregulation of DGKε perturbs lipid signali
Tue Mar 07 00:00:00 UTC 2017
A new method for post-translationally labeling proteins in live cells for fluorescence imaging and tracking.
Protein engineering, design & selection : PEDS
Hinrichsen M,Lenz M,Edwards JM,Miller OK,Mochrie SGJ,Swain PS,Schwarz-Linek U,Regan L
We present a novel method to fluorescently label proteins, post-translationally, within live Saccharomycescerevisiae. The premise underlying this work is that fluorescent protein (FP) tags are less disruptive to normal processing and function when they are attached post-translationally, because target proteins are allowed to fold properly and reach their final subcellular location before being labeled. We accomplish this post-translational labeling by expressing the target protein fused to a sho
Fri Dec 01 00:00:00 UTC 2017
CBMs as Probes to Explore Plant Cell Wall Heterogeneity Using Immunocytochemistry.
Methods in molecular biology (Clifton, N.J.)
Badruna L,Burlat V,Montanier CY
Immunocytochemistry is a widely used technique to localize antigen within intact tissues. Plant cell walls are complex matrixes of highly decorated polysaccharides and the large number of CBM families displaying specific substrate recognition reflects this complexity. The accessibility of large proteins, such as antibodies, to their cell wall epitopes may be sometimes difficult due to steric hindrance problems. Due to their smaller size, CBMs are interesting alternative probes. The aim of this c
Fri Apr 21 00:00:00 UTC 2017
FGF1 protects neuroblastoma SH-SY5Y cells from p53-dependent apoptosis through an intracrine pathway regulated by FGF1 phosphorylation.
Cell death & disease
Pirou C,Montazer-Torbati F,Jah N,Delmas E,Lasbleiz C,Mignotte B,Renaud F
Neuroblastoma a sympathetic nervous system tumor accounts for 15% of cancer deaths in children. In contrast to most human tumors p53 is rarely mutated in human primary neuroblastoma suggesting impaired p53 activation in neuroblastoma. Various studies have shown correlations between fgf1 expression levels and both prognosis severity and tumor chemoresistance. As we previously showed that fibroblast growth factor 1 (FGF1) inhibited p53-dependent apoptosis in neuron-like PC12 cells we initiated the
Thu Aug 31 00:00:00 UTC 2017
Blue light and CO2 signals converge to regulate light-induced stomatal opening.
Nature communications
Hiyama A,Takemiya A,Munemasa S,Okuma E,Sugiyama N,Tada Y,Murata Y,Shimazaki KI
Stomata regulate gas exchange between plants and atmosphere by integrating opening and closing signals. Stomata open in response to low CO concentrations to maximize photosynthesis in the light; however the mechanisms that coordinate photosynthesis and stomatal conductance have yet to be identified. Here we identify and characterize CBC1/2 (CONVERGENCE OF?BLUE LIGHT (BL) AND CO 1/2) two kinases that link BL a major component of photosynthetically active radiation (PAR) and the signals from low c
Fri Nov 03 00:00:00 UTC 2017
TALE proteins search DNA using a rotationally decoupled mechanism.
Nature chemical biology
Cuculis L,Abil Z,Zhao H,Schroeder CM
Transcription activator-like effector (TALE) proteins are a class of programmable DNA-binding proteins used extensively for gene editing. Despite recent progress, however, little is known about their sequence search mechanism. Here, we use single-molecule experiments to study TALE search along DNA. Our results show that TALEs utilize a rotationally decoupled mechanism for nonspecific search, despite remaining associated with DNA templates during the search process. Our results suggest that the p
Sat Oct 01 00:00:00 UTC 2016
Functional protein-based nanomaterial produced in microorganisms recognized as safe: A new platform for biotechnology.
Acta biomaterialia
Cano-Garrido O,Sánchez-Chardi A,Parés S,Giró I,Tatkiewicz WI,Ferrer-Miralles N,Ratera I,Natalello A,Cubarsi R,Veciana J,Bach À,Villaverde A,Arís A,Garcia-Fruitós E
Inclusion bodies (IBs) are protein-based nanoparticles formed in Escherichia coli through stereospecific aggregation processes during the overexpression of recombinant proteins. In the last years, it has been shown that IBs can be used as nanostructured biomaterials to stimulate mammalian cell attachment, proliferation, and differentiation. In addition, these nanoparticles have also been explored as natural delivery systems for protein replacement therapies. Although the production of these prot
Sat Oct 01 00:00:00 UTC 2016
RAD18, WRNIP1 and ATMIN promote ATM signalling in response to replication stress.
Oncogene
Kanu N,Zhang T,Burrell RA,Chakraborty A,Cronshaw J,DaCosta C,Grönroos E,Pemberton HN,Anderton E,Gonzalez L,Sabbioneda S,Ulrich HD,Swanton C,Behrens A
The DNA replication machinery invariably encounters obstacles that slow replication fork progression, and threaten to prevent complete replication and faithful segregation of sister chromatids. The resulting replication stress activates ATR, the major kinase involved in resolving impaired DNA replication. In addition, replication stress also activates the related kinase ATM, which is required to prevent mitotic segregation errors. However, the molecular mechanism of ATM activation by replication
Thu Jul 28 00:00:00 UTC 2016
An unrecognized function for COPII components in recruiting the viral replication protein BMV 1a to the perinuclear ER.
Journal of cell science
Li J,Fuchs S,Zhang J,Wellford S,Schuldiner M,Wang X
Positive-strand RNA viruses invariably assemble their viral replication complexes (VRCs) by remodeling host intracellular membranes. How viral replication proteins are targeted to specific organelle membranes to initiate VRC assembly remains elusive. Brome mosaic virus (BMV), whose replication can be recapitulated in Saccharomyces cerevisiae, assembles its VRCs by invaginating the outer perinuclear endoplasmic reticulum (ER) membrane. Remarkably, BMV replication protein 1a (BMV 1a) is the only v
Sat Oct 01 00:00:00 UTC 2016
Expression of sheep pathogen Babesia sp. Xinjiang rhoptry-associated protein 1 and evaluation of its diagnostic potential by enzyme-linked immunosorbent assay.
Parasitology
Niu Q,Liu Z,Yang J,Yu P,Pan Y,Zhai B,Luo J,Guan G,Yin H
Ovine babesiosis is one of the most important tick-borne haemoparasitic diseases of small ruminants. The ovine parasite Babesia sp. Xinjiang is widespread in China. In this study, recombinant full-length XJrRAP-1aα2 (rhoptry-associated protein 1aα2) and C-terminal XJrRAP-1aα2 CT of Babesia sp. Xinjiang were expressed and used to evaluate their diagnostic potential for Babesia sp. Xinjiang infections by indirect enzyme-linked immunosorbent assay (ELISA). Purified XJrRAP-1a&alpha
Thu Dec 01 00:00:00 UTC 2016
Conserved transmembrane glycine residues in the Shigella flexneri polysaccharide co-polymerase protein WzzB influence protein-protein interactions.
Microbiology (Reading, England)
Papadopoulos M,Tran ENH,Murray GL,Morona R
The O antigen (Oag) component of lipopolysaccharides (LPS) is crucial for virulence and Oag chain-length regulation is controlled by the polysaccharide co-polymerase class 1 (PCP1) proteins. Crystal structure analyses indicate that structural conservation among PCP1 proteins is highly maintained, however the mechanism of Oag modal-chain-length control remains to be fully elucidated. Shigella flexneri PCP1 protein WzzBSF confers a modal-chain length of 10-17 Oag repeat units (RUs), whereas the Sa
Wed Jun 01 00:00:00 UTC 2016
Identification of an epitope within the Bovine herpesvirus 1 glycoprotein E cytoplasmic tail and use of a monoclonal antibody directed against the epitope for the differentiation between vaccinated and infected animals.
Journal of virological methods
Chowdhury SI
We constructed a recombinant bovine herpesvirus type 1 triple mutant virus (BoHV-1 tmv) that lacks UL49.5 residues 30-32 and 80-96, gE cytoplasmic tail (gE CT) residues 452-575 and the entire 435 bp long Us9 ORF. To develop a gE CT-specific blocking ELISA test that is necessary to distinguish the BoHV-1 tmv vaccinated calves from the wild-type (wt) virus-infected calves, a mouse monoclonal antibody (mAb) 2H8F3 was generated by using the Escherichia coli expressed gE CT residues 452-575. Further,
Fri Jul 01 00:00:00 UTC 2016
DEEPN as an Approach for Batch Processing of Yeast 2-Hybrid Interactions.
Cell reports
Pashkova N,Peterson TA,Krishnamani V,Breheny P,Stamnes M,Piper RC
We adapted the yeast 2-hybrid assay to simultaneously uncover multiple transient protein interactions within a single screen by using a strategy termed DEEPN (dynamic enrichment for evaluation of protein networks). This approach incorporates high-throughput DNA sequencing and computation to follow competition among a plasmid population encoding interacting partners. To demonstrate the capacity of DEEPN, we identify a wide range of ubiquitin-binding proteins, including interactors that we verify
Tue Sep 27 00:00:00 UTC 2016
Stereochemical inversion of (S)-reticuline by a cytochrome P450 fusion in opium poppy.
Nature chemical biology
Farrow SC,Hagel JM,Beaudoin GA,Burns DC,Facchini PJ
The gateway to morphine biosynthesis in opium poppy (Papaver somniferum) is the stereochemical inversion of (S)-reticuline since the enzyme yielding the first committed intermediate salutaridine is specific for (R)-reticuline. A fusion between a cytochrome P450 (CYP) and an aldo-keto reductase (AKR) catalyzes the S-to-R epimerization of reticuline via 12-dehydroreticuline. The reticuline epimerase (REPI) fusion was detected in opium poppy and in Papaver bracteatum which accumulates thebaine. In
Tue Sep 01 00:00:00 UTC 2015
A new cell-penetrating peptide that blocks the autoinhibitory XIP domain of NCX1 and enhances antiporter activity.
Molecular therapy : the journal of the American Society of Gene Therapy
Molinaro P,Pannaccione A,Sisalli MJ,Secondo A,Cuomo O,Sirabella R,Cantile M,Ciccone R,Scorziello A,di Renzo G,Annunziato L
The plasma membrane Na(+)/Ca(2+) exchanger (NCX) is a high-capacity ionic transporter that exchanges 3Na(+) ions for 1Ca(2+) ion. The first 20 amino acids of the f-loop named exchanger inhibitory peptide (XIP(NCX1)) represent an autoinhibitory region involved in the Na(+)-dependent inactivation of the exchanger. Previous research has shown that an exogenous peptide having the same amino acid sequence as the XIP(NCX1) region exerts an inhibitory effect on NCX activity. In this study we identified
Sun Mar 01 00:00:00 UTC 2015
Endogenous IGFBP-3 Mediates Intrinsic Apoptosis Through Modulation of Nur77 Phosphorylation and Nuclear Export.
Endocrinology
Agostini-Dreyer A,Jetzt AE,Stires H,Cohick WS
In nontransformed bovine mammary epithelial cells, the intrinsic apoptosis inducer anisomycin (ANS) induces IGFBP-3 expression and nuclear localization and knockdown of IGFBP-3 attenuates ANS-induced apoptosis. Others have shown in prostate cancer cells that exogenous IGFBP-3 induces apoptosis by facilitating nuclear export of the orphan nuclear receptor Nur77 and its binding partner, retinoid X receptor-α (RXRα). The goal of the present work was to determine whether endogenous IGFBP
Sun Nov 01 00:00:00 UTC 2015
Phosphorylated and unphosphorylated serine 13 of CDC37 stabilize distinct interactions between its client and HSP90 binding domains.
Biochemistry
Liu W,Landgraf R
Folding and maturation of most protein kinases require chaperone assistance. In higher eukaryotes, CDC37 is the predominant cochaperone that facilitates the transfer of kinase clients to HSP90. Kinase recognition is thought to occur through the N-terminal domain, which has, thus far, eluded structure determination. Client processing also requires the phosphorylation of the N-terminal tail at Ser13 by protein kinase CK2 (casein kinase 2). How phosphorylation alters the molecular properties of CDC
Tue Feb 24 00:00:00 UTC 2015
Immunocapture-Selected Reaction Monitoring Screening Facilitates the Development of ELISA for the Measurement of Native TEX101 in Biological Fluids.
Molecular & cellular proteomics : MCP
Korbakis D,Brinc D,Schiza C,Soosaipillai A,Jarvi K,Drabovich AP,Diamandis EP
Monoclonal antibodies that bind the native conformation of proteins are indispensable reagents for the development of immunoassays, production of therapeutic antibodies and delineating protein interaction networks by affinity purification-mass spectrometry. Antibodies generated against short peptides, protein fragments or even full length recombinant proteins may not bind the native protein form in biological fluids, thus limiting their utility. Here, we report the application of immunocapture c
Mon Jun 01 00:00:00 UTC 2015
Bimodal activation of BubR1 by Bub3 sustains mitotic checkpoint signaling.
Proceedings of the National Academy of Sciences of the United States of America
Han JS,Vitre B,Fachinetti D,Cleveland DW
The mitotic checkpoint (also known as the spindle assembly checkpoint) prevents premature anaphase onset through generation of an inhibitor of the E3 ubiquitin ligase APC/C, whose ubiquitination of cyclin B and securin targets them for degradation. Combining in vitro reconstitution and cell-based assays, we now identify dual mechanisms through which Bub3 promotes mitotic checkpoint signaling. Bub3 enhances signaling at unattached kinetochores not only by facilitating binding of BubR1 but also by
Tue Oct 07 00:00:00 UTC 2014
Lys11- and Lys48-linked ubiquitin chains interact with p97 during endoplasmic-reticulum-associated degradation.
The Biochemical journal
Locke M,Toth JI,Petroski MD
The ATPase associated with various cellular activities p97 has a critical function in the cytoplasmic degradation of proteins misfolded in the ER (endoplasmic reticulum) through a mechanism known as ERAD (ER-associated degradation). During this process, p97 binds polyubiquitinated ERAD substrates and couples ATP hydrolysis to their dislocation from the ER as a prerequisite to destruction by the proteasome. The ubiquitin signals important for this process are not fully understood. In the present
Tue Apr 01 00:00:00 UTC 2014
Delineation of domains within the cannabinoid CB1 and dopamine D2 receptors that mediate the formation of the heterodimer complex.
Journal of molecular neuroscience : MN
Khan SS,Lee FJ
Both the cannabinoid CB1 receptor (CB1) and dopamine D2 receptor (D2R) are G protein-coupled receptors that are linked to inhibitory Gαi/o protein, whereby activation of the receptor leads to the inhibition of cAMP production. Moreover, previous findings have shown evidence of cross-talk between the dopamine and endocannabinoid systems. In this report, we confirm the interaction of CB1 and D2R with co-immunoprecipitation experiments using human embryonic kidney 293T (HEK-293T) cells co-exp
Thu May 01 00:00:00 UTC 2014
An RNA degradation machine sculpted by Ro autoantigen and noncoding RNA.
Cell
Chen X,Taylor DW,Fowler CC,Galan JE,Wang HW,Wolin SL
SummaryMany bacteria contain an ortholog of the Ro autoantigen, a ring-shaped protein that binds noncoding RNAs (ncRNAs) called Y RNAs. In the only studied bacterium, Deinococcus radiodurans, the Ro ortholog Rsr functions in heat-stress-induced ribosomal RNA (rRNA) maturation and starvation-induced rRNA decay. However, the mechanism by which this conserved protein and its associated ncRNAs act has been obscure. We report that Rsr and the exoribonuclease polynucleotide phosphorylase (PNPase) form
Thu Mar 28 00:00:00 UTC 2013
Catalytic assembly of the mitotic checkpoint inhibitor BubR1-Cdc20 by a Mad2-induced functional switch in Cdc20.
Molecular cell
Han JS,Holland AJ,Fachinetti D,Kulukian A,Cetin B,Cleveland DW
SummaryThe mitotic checkpoint acts to maintain chromosome content by generation of a diffusible anaphase inhibitor. Unattached kinetochores catalyze a conformational shift in Mad2, converting an inactive open form into a closed form that can capture Cdc20, the mitotic activator of the APC/C ubiquitin ligase. Mad2 binding is now shown to promote a functional switch in Cdc20, exposing a previously inaccessible site for binding to BubR1’s conserved Mad3 homology domain. BubR1, but not Ma
Thu Jul 11 00:00:00 UTC 2013
Nucleosome-binding activities within JARID2 and EZH1 regulate the function of PRC2 on chromatin.
Genes & development
Son J,Shen SS,Margueron R,Reinberg D
Polycomb-repressive complex 2 (PRC2) comprises specific members of the Polycomb group of epigenetic modulators. PRC2 catalyzes methylation of histone H3 at Lys 27 (H3K27me3) through its Enhancer of zeste (Ezh) constituent, of which there are two mammalian homologs: Ezh1 and Ezh2. Several ancillary factors, including Jarid2, modulate PRC2 function, with Jarid2 facilitating its recruitment to target genes. Jarid2, like Ezh2, is present in poorly differentiated and actively dividing cells, while Ez
Sun Dec 15 00:00:00 UTC 2013
Structural basis for endosomal trafficking of diverse transmembrane cargos by PX-FERM proteins.
Proceedings of the National Academy of Sciences of the United States of America
Ghai R,Bugarcic A,Liu H,Norwood SJ,Skeldal S,Coulson EJ,Li SS,Teasdale RD,Collins BM
Transit of proteins through the endosomal organelle following endocytosis is critical for regulating the homeostasis of cell-surface proteins and controlling signal transduction pathways. However, the mechanisms that control these membrane-transport processes are poorly understood. The Phox-homology (PX) domain-containing proteins sorting nexin (SNX) 17, SNX27, and SNX31 have emerged recently as key regulators of endosomal recycling and bind conserved Asn-Pro-Xaa-Tyr-sorting signals in transmemb
Tue Feb 19 00:00:00 UTC 2013
Npas4 is activated by melatonin, and drives the clock gene Cry1 in the ovine pars tuberalis.
Molecular endocrinology (Baltimore, Md.)
West A,Dupré SM,Yu L,Paton IR,Miedzinska K,McNeilly AS,Davis JR,Burt DW,Loudon AS
Seasonal mammals integrate changes in the duration of nocturnal melatonin secretion to drive annual physiologic cycles. Melatonin receptors within the proximal pituitary region, the pars tuberalis (PT), are essential in regulating seasonal neuroendocrine responses. In the ovine PT, melatonin is known to influence acute changes in transcriptional dynamics coupled to the onset (dusk) and offset (dawn) of melatonin secretion, leading to a potential interval-timing mechanism capable of decoding chan
Sat Jun 01 00:00:00 UTC 2013
A strategy for the expression of recombinant proteins traditionally hard to purify.
Analytical biochemistry
Frank EG,McDonald JP,Karata K,Huston D,Woodgate R
We have developed a series of plasmid vectors for the soluble expression and subsequent purification of recombinant proteins that have historically proven to be extremely difficult to purify from Escherichia coli. Instead of dramatically overproducing the target protein, it is expressed at a low basal level that facilitates the correct folding of the recombinant protein and increases its solubility. Highly active recombinant proteins that are traditionally difficult to purify are readily purifie
Mon Oct 15 00:00:00 UTC 2012
Intranasal delivery of caspase-9 inhibitor reduces caspase-6-dependent axon/neuron loss and improves neurological function after stroke.
The Journal of neuroscience : the official journal of the Society for Neuroscience
Akpan N,Serrano-Saiz E,Zacharia BE,Otten ML,Ducruet AF,Snipas SJ,Liu W,Velloza J,Cohen G,Sosunov SA,Frey WH,Salvesen GS,Connolly ES,Troy CM
Despite extensive research to develop an effective neuroprotective strategy for the treatment of ischemic stroke, therapeutic options remain limited. Although caspase-dependent death is thought to play a prominent role in neuronal injury, direct evidence of active initiator caspases in stroke and the functional relevance of this activity have not previously been shown. Using an unbiased caspase-trapping technique in vivo, we isolated active caspase-9 from ischemic rat brain within 1 h of reperfu
Wed Jun 15 00:00:00 UTC 2011
Identification of the atypical MAPK Erk3 as a novel substrate for p21-activated kinase (Pak) activity.
The Journal of biological chemistry
De la Mota-Peynado A,Chernoff J,Beeser A
The class I p21-activated kinases (Pak1-3) regulate many essential biological processes, including cytoskeletal rearrangement, cell cycle progression, apoptosis, and cellular transformation. Although many Pak substrates, including elements of MAPK signaling cascades, have been identified, it is likely that additional substrates remain to be discovered. Identification of such substrates, and determination of the consequences of their phosphorylation, is essential for a better understanding of cla
Fri Apr 15 00:00:00 UTC 2011
The transmembrane domain 6 of vacuolar H(+)-pyrophosphatase mediates protein targeting and proton transport.
Biochimica et biophysica acta
Pan YJ,Lee CH,Hsu SH,Huang YT,Lee CH,Liu TH,Chen YW,Lin SM,Pan RL
Vacuolar H+-pyrophosphatase (V-PPase; EC 3.6.1.1) plays a significant role in the maintenance of the pH in cytoplasm and vacuoles via proton translocation from the cytosol to the vacuolar lumen at the expense of PPi hydrolysis. The topology of V-PPase as predicted by TopPred II suggests that the catalytic site is putatively located in loop e and exposed to the cytosol. The adjacent transmembrane domain 6 (TM6) is highly conserved and believed to participate in the catalytic function and conforma
Sat Jan 01 00:00:00 UTC 2011
The transcription factor MafB antagonizes antiviral responses by blocking recruitment of coactivators to the transcription factor IRF3.
Nature immunology
Kim H,Seed B
Viral infection induces type I interferons (IFN-alpha and IFN-beta) that recruit unexposed cells in a self-amplifying response. We report that the transcription factor MafB thwarts auto-amplification by a metastable switch activity. MafB acted as a weak positive basal regulator of transcription at the IFNB1 promoter through activity at transcription factor AP-1-like sites. Interferon elicitors recruited the transcription factor IRF3 to the promoter, whereupon MafB acted as a transcriptional anta
Sun Aug 01 00:00:00 UTC 2010
Localization of type-2 angiotensin II receptor in adrenal gland.
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
Harada K,Matsuoka H,Fujimoto N,Endo Y,Hasegawa Y,Matsuo A,Kikuchi Y,Matsumoto T,Inoue M
The localization of the type-2 angiotensin II receptor (AT2) in the adrenal glands of rats, guinea pigs, bovines, and humans was examined at the mRNA and protein levels. PCR products for AT2 were detected in the adrenal cortices and adrenal medullae of all the mammals examined with an RT-PCR technique. Three different anti-AT2 antibodies (Abs), whose specificity was confirmed in our hands, recognized a 50-kDa protein in the adrenal glands of the four mammals, and this recognition was abolished b
Thu Jul 01 00:00:00 UTC 2010
A specific docking site for DNA polymerase {alpha}-primase on the SV40 helicase is required for viral primosome activity, but helicase activity is dispensable.
The Journal of biological chemistry
Huang H,Zhao K,Arnett DR,Fanning E
Replication of simian virus 40 (SV40) DNA, a model for eukaryotic chromosomal replication, can be reconstituted in vitro using the viral helicase (large tumor antigen, or Tag) and purified human proteins. Tag interacts physically with two cellular proteins, replication protein A and DNA polymerase α-primase (pol-prim), constituting the viral primosome. Like the well characterized primosomes of phages T7 and T4, this trio of proteins coordinates parental DNA unwinding with primer synthesis
Fri Oct 22 00:00:00 UTC 2010
