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Journal of Molecular Biology

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https://www.readbyqxmd.com/read/28822683/hsp90-sensitivity-to-adp-reveals-hidden-regulation-mechanisms
#1
Jackson C Halpin, Timothy O Street
The ATPase cycle of the Hsp90 molecular chaperone is essential for maintaining the stability of numerous client proteins. Extensive analysis has focused on ATP-driven conformational changes of Hsp90, however, little is known about how Hsp90 operates under physiological nucleotide conditions in which both ATP and ADP are present. By quantifying Hsp90 activity under mixed nucleotide conditions we find dramatic differences in ADP-sensitivity among Hsp90 homologs. ADP acts as a strong ATPase inhibitor of cytosol-specific Hsp90 homologs, whereas organellular Hsp90 homologs (Grp94 and TRAP1) are relatively insensitive to the presence of ADP...
August 16, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28821455/rnase-hii-saves-rnha-mutant-escherichia-coli-from-r-loop-associated-chromosomal-fragmentation
#2
Elena A Kouzminova, Farid F Kadyrov, Andrei Kuzminov
The rnhAB mutant E. coli, deficient in two RNase H enzymes that remove both R-loops and incorporated ribonucleotides (rNs) from DNA, grow slowly, suggesting accumulation of rN-containing DNA lesions (R-lesions). We report that the rnhAB mutants have reduced viability, form filaments with abnormal nucleoids, induce SOS and fragment their chromosome, revealing replication and/or segregation stress. R-loops are known to interfere with replication forks, and sensitivity of the double rnhAB mutants to translation inhibition points to R-loops as precursors for R-lesions...
August 15, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28818634/mining-na%C3%A3-ve-rabbit-antibody-repertoires-by-phage-display-for-monoclonal-antibodies-of-therapeutic-utility
#3
Haiyong Peng, Thomas Nerreter, Jing Chang, Junpeng Qi, Xiuling Li, Pabalu Karunadharma, Gustavo Martinez, Mohammad Fallahi, Jo Soden, Jim Freeth, Roger R Beerli, Ulf Grawunder, Michael Hudecek, Christoph Rader
Owing to their high affinities and specificities, rabbit monoclonal antibodies (mAbs) have demonstrated value and potential primarily as basic research and diagnostic reagents, but in some cases also as therapeutics. To accelerate access to rabbit mAbs bypassing immunization, we generated a large naïve rabbit antibody repertoire represented by a phage display library encompassing >10 billion independent antibodies in chimeric rabbit/human Fab format and validated it by next-generation sequencing. Panels of rabbit mAbs selected from this library against two emerging cancer targets, ROR1 and ROR2, revealed high diversity, affinity, and specificity...
August 14, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28807855/spatial-organization-and-molecular-interactions-of-the-schizosaccharomyces-pombe-ccq1-tpz1-poz1-shelterin-complex
#4
Harry Scott, Jin-Kwang Kim, Clinton Yu, Lan Huang, Feng Qiao, Derek J Taylor
The shelterin complex is a macromolecular assembly of proteins that binds to and protects telomeric DNA, which composes the ends of all linear chromosomes. Shelterin proteins prevent chromosome ends from fusing together and from eliciting erroneous induction of DNA damage response pathways. In addition, shelterin proteins play key roles in regulating the recruitment and activation of telomerase, an enzyme that extends telomeric DNA. In fission yeast, Schizosaccharomyces pombe, interactions between the shelterin proteins Ccq1, Tpz1, and Poz1 are important for regulating telomerase-mediated telomere synthesis and thus telomere length homeostasis...
August 12, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28802871/stack-locally-and-act-globally-a-few-nucleotides-make-all-the-difference-in-enterovirus-71-ires-binding-hnrnap-a1-and-infectious-phenotypes-commentary-on-hnrnp-a1-alters-the-structure-of-a-conserved-enterovirus-ires-domain-to-stimulate-viral-translation
#5
https://www.readbyqxmd.com/read/28782559/trap-seq-an-rna-sequencing-based-pipeline-for-the-identification-of-gene-trap-insertions-in-mammalian-cells
#6
Cristina Mayor-Ruiz, Orlando Dominguez, Oscar Fernandez-Capetillo
The development of haploid mammalian cell lines, coupled to next-generation sequencing, has recently facilitated forward genetic screenings in mammals. For mutagenesis, retrovirus- or transposon-based gene traps are frequently used. Current methods to map gene-trap insertions are based on inverse or splinkerette PCR, which despite their efficacy are prone to artifacts and do not provide information on expression of the targeted gene. Here, we describe a new RNA sequencing-based method (Trap(Seq)) to map gene-trap insertions...
August 4, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28782560/glutamate-promotes-ssb-protein-protein-interactions-via-intrinsically-disordered-regions
#7
Alexander G Kozlov, Min Kyung Shinn, Elizabeth A Weiland, Timothy M Lohman
E. coli single strand (ss) DNA binding protein (SSB) is an essential protein that binds to ssDNA intermediates formed during genome maintenance. SSB homotetramers bind ssDNA in several modes that differ in occluded site size and cooperativity. High "unlimited" cooperativity is associated with the 35 site size ((SSB)35) mode at low [NaCl], whereas the 65 site size ((SSB)65) mode formed at higher [NaCl] (> 200mM), where ssDNA wraps completely around the tetramer, displays "limited" cooperativity forming dimers of tetramers...
August 3, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28782558/exploring-the-denatured-state-ensemble-by-single-molecule-chemo-mechanical-unfolding-the-effect-of-force-temperature-and-urea
#8
Emily Guinn, Susan Marqusee
While it is widely appreciated that the denatured state of a protein is a heterogeneous conformational ensemble, there is still debate over how this ensemble changes with environmental conditions. Here, we use single-molecule chemo-mechanical unfolding, which combines force and urea using the optical tweezers, together with traditional protein unfolding studies to explore how perturbants commonly used to unfold proteins (urea, force and temperature) affect the denatured state ensemble. We compare the urea m-values, which report on the change in solvent accessible surface area for unfolding, to probe the denatured state as a function of force, temperature and urea...
August 3, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28760452/marking-the-milestones-in-structural-biology
#9
EDITORIAL
Peter E Wright, Hélène Hodak
No abstract text is available yet for this article.
July 29, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28760365/corrigendum-to-mir-125a-5p-modulates-phenotypic-switch-of-vascular-smooth-muscle-cells-by-targeting-ets-1-j-mol-biol-jun-16-2017-429-12-1817-1828
#10
C Gareri, C Iaconetti, S Sorrentino, C Covello, S De Rosa, C Indolfi
No abstract text is available yet for this article.
July 28, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28757069/structural-foundations-of-rna-silencing-by-argonaute
#11
REVIEW
Jessica Sheu-Gruttadauria, Ian J MacRae
Nearly every cell in the human body contains a set of programmable gene-silencing proteins named Argonaute. Argonaute proteins mediate gene regulation by small RNAs and thereby contribute to cellular homeostasis during diverse physiological process, such as stem cell maintenance, fertilization, and heart development. Over the last decade, remarkable progress has been made toward understanding Argonaute proteins, small RNAs, and their roles in eukaryotic biology. Here, we review current understanding of Argonaute proteins from a structural prospective and discuss unanswered questions surrounding this fascinating class of enzymes...
July 27, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28756087/a-glimpse-of-membrane-transport-through-structures-advances-in-the-structural-biology-of-the-glut-glucose-transporters
#12
REVIEW
Nieng Yan
The cellular uptake of glucose is an essential physiological process, and movement of glucose across biological membranes requires specialized transporters. The major facilitator superfamily glucose transporters GLUTs, encoded by the SLC2A genes, have been a paradigm for functional, mechanistic, and structural understanding of solute transport in the past century. This review starts with a glimpse into the structural biology of membrane proteins and particularly membrane transport proteins, enumerating the landmark structures in the past 25years...
July 26, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28754374/oh1-from-orf-virus-a-new-tyrosine-phosphatase-that-displays-distinct-structural-features-and-triple-substrate-specificity
#13
Danilo Segovia, Ahmed Haouz, Darío Porley, Natalia Olivero, Mariano Martínez, Mahendra Mariadassou, Mabel Berois, Gwenaëlle André-Leroux, Andrea Villarino
Viral tyrosine phosphatases such as VH1 from Vaccinia and Variola virus are recognized as important effectors of host-pathogen interactions. While proteins sharing sequence to VH1 have been identified in other viruses, their structural and functional characterization is not known. In this work, we determined the crystal structure of the VH1 homologue in the Orf virus, herein named OH1. Similarly to Variola and Vaccinia VH1, the structure of OH1 shows a dimer with the typical dual specificity phosphatase fold...
July 25, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28743590/sequence-specificity-in-the-entropy-driven-binding-of-a-small-molecule-and-a-disordered-peptide
#14
Gabriella T Heller, Francesco A Aprile, Massimiliano Bonomi, Carlo Camilloni, Alfonso De Simone, Michele Vendruscolo
Approximately one-third of the human proteome is made up of proteins that are entirely disordered or that contain extended disordered regions. Although these disordered proteins are closely linked with many major diseases, their binding mechanisms with small molecules remain poorly understood, and a major concern is whether their specificity can be sufficient for drug development. Here, by studying the interaction of a small molecule and a disordered peptide from the oncogene protein c-Myc, we describe a "specific-diffuse" binding mechanism that exhibits sequence specificity despite being of entropic nature...
July 22, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28739480/the-changing-face-of-aging-highly-sulfated-glycosaminoglycans-induce-amyloid-formation-in-a-lattice-corneal-dystrophy-model-protein
#15
Kirsten G Malmos, Marcel Stenvang, Cagla Sahin, Gunna Christiansen, Daniel Otzen
Glycosaminoglycans (GAGs) are related to multiple biological functions and diseases. There is growing evidence that GAG concentration and sulfate content increase with age. The destabilizing mutation A546T in the corneal protein TGFBIp leads to lattice-type corneal dystrophy, but symptoms only appear in the fourth decade of life. We hypothesize that this delayed phenotype can be explained by increased GAG sulfation over time. Using in vitro assays with the C-terminal TGFIBIp domain Fas1-4, previously shown to recapitulate many properties of full-length TGFBIp, we find that only long GAGs with multiple sulfate groups on each repeating unit increase the amount of worm-like aggregates and induce long, straight fibrils in A546T...
July 21, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28736176/crystal-structure-of-the-c-terminal-domain-of-human-eif2d-and-its-implications-on-eukaryotic-translation-initiation
#16
Anand T Vaidya, Ivan B Lomakin, Newlyn N Joseph, Sergey E Dmitriev, Thomas A Steitz
Protein synthesis is a key process in all living organisms. In eukaryotes, Initiation Factor 2 (eIF2) plays an important role in translation initiation as it selects and delivers the initiator tRNA to the small ribosomal subunit. Under stress conditions, phosphorylation of the α-subunit of eIF2 downregulates cellular protein synthesis. However, translation of certain mRNAs continues via the eIF2D dependent non-canonical initiation pathway. The molecular mechanism of this process remains elusive. In addition, eIF2D plays a role in translation re-initiation and ribosome recycling...
July 20, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28736175/structural-basis-for-receptor-selectivity-by-the-whitewater-arroyo-mammarenavirus
#17
Amir Shimon, Orly Shani, Ron Diskin
Whitewater Arroyo Virus belongs to the 'New World' group of mammarenaviruses, which reside in rodent reservoirs and are prevalent in North and South Americas. Clades B and A/B of New World mammarenaviruses use transferrin receptor 1 (TfR1) for entry. While all of these viruses use rodent-derived TfR1 orthologs, some can also use the human-TfR1 and thereby infect humans. Although we have structural information for TfR1 recognition by pathogenic virus, we do not know what the structural differences are between the receptor binding domains of pathogenic and non-pathogenic viruses that allow some but not all viruses to utilize the human receptor for entry...
July 20, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28733144/the-spliceosome-a-protein-directed-metalloribozyme
#18
REVIEW
Yigong Shi
Pre-mRNA splicing is executed by the ribonucleoprotein machinery spliceosome. Nearly 40 years after the discovery of pre-mRNA splicing, the atomic structure of the spliceosome has finally come to light. Four distinct conformational states of the yeast spliceosome have been captured at atomic or near-atomic resolutions. Two catalytic metal ions at the active site are specifically coordinated by the U6 small nuclear RNA (snRNA) and catalyze both the branching reaction and the exon ligation. Of the three snRNAs in the fully assembled spliceosome, U5 and U6, along with 30 contiguous nucleotides of U2 at its 5'-end, remain structurally rigid throughout the splicing reaction...
July 19, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28733143/mechanisms-of-nlrp1-mediated-autoinflammatory-disease-in-humans-and-mice
#19
REVIEW
Chien-Hsiung Yu, Jonas Moecking, Matthias Geyer, Seth L Masters
NLRP1 was the first NOD-like receptor described to form an inflammasome, recruiting ASC to activate caspase-1, which processes interleukin-1β and interleukin-18 to their active form. A wealth of new genetic information has now redefined our understanding of this innate immune sensor. Specifically, rare loss-of-function variants in the N-terminal pyrin domain indicate that this part of NLRP1 is autoinhibitory and normally acts to prevent a familial autoinflammatory skin disease associated with cancer. In the absence of a ligand to trigger human NLRP1, these mutations have now confirmed the requirement of NLRP1 autolytic cleavage within the FIIND domain, which had previously been implicated in NLRP1 activation...
July 19, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28728980/the-role-of-hexon-protein-as-a-molecular-mold-in-patterning-the-protein-ix-organization-in-human-adenoviruses
#20
Vijay S Reddy
Adenoviruses are respiratory, ocular and enteric pathogens that form complex capsids, which are assembled from seven different structural proteins and composed of several core proteins that closely interact with the packaged dsDNA genome. The recent near-atomic resolution structures revealed that the interlacing continuous hexagonal network formed by the protein IX molecules is conserved among different human adenoviruses (HAdVs), but not in non-HAdVs. In this report, we propose a distinct role for the hexon protein as a "molecular mold" in enabling the formation of such hexagonal protein IX network that has been shown to preserve the stability and infectivity of HAdVs...
July 18, 2017: Journal of Molecular Biology
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