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The inhibition of cell proliferation by ouabain has been analyzed with respect to the cell cycle. Three lines of evidence indicate that growth rate is modified by altering to different degrees the rate of progress through stages of the cell cycle: (1) a three hour lag occurs between the time of ouabain addition and the inhibition of proliferation; (2) ouabain must be present at least two to four hours prior to the mitotic burst of synchronized cells for inhibition of mitosis to occur; (3) parasynchrony is observed when cells are resuspended in ouabain-free medium after 12 hours of exposure to ouabain. Analysis of the distribution of cells in each of the stages of the cell cycle at various times during ouabain treatment reveals a progressive increase in the fraction of cells in S with a concomitant decrease in the percent of cells in each of the other stages. These results indicate that the prolongation of the cell cycle time in the presence of ouabain is due primarily to an S stage block. 相似文献
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Jeremy R. Lohman Ming Ma Marianne E. Cuff Lance Bigelow Jessica Bearden Gyorgy Babnigg Andrzej Joachimiak George N. Phillips Jr. Ben Shen 《Proteins》2014,82(7):1210-1218
Carrier proteins (CPs) play a critical role in the biosynthesis of various natural products, especially in nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) enzymology, where the CPs are referred to as peptidyl‐carrier proteins (PCPs) or acyl‐carrier proteins (ACPs), respectively. CPs can either be a domain in large multifunctional polypeptides or standalone proteins, termed Type I and Type II, respectively. There have been many biochemical studies of the Type I PKS and NRPS CPs, and of Type II ACPs. However, recently a number of Type II PCPs have been found and biochemically characterized. In order to understand the possible interaction surfaces for combinatorial biosynthetic efforts we crystallized the first characterized and representative Type II PCP member, BlmI, from the bleomycin biosynthetic pathway from Streptomyces verticillus ATCC 15003. The structure is similar to CPs in general but most closely resembles PCPs. Comparisons with previously determined PCP structures in complex with catalytic domains reveals a common interaction surface. This surface is highly variable in charge and shape, which likely confers specificity for interactions. Previous nuclear magnetic resonance (NMR) analysis of a prototypical Type I PCP excised from the multimodular context revealed three conformational states. Comparison of the states with the structure of BlmI and other PCPs reveals that only one of the NMR states is found in other studies, suggesting the other two states may not be relevant. The state represented by the BlmI crystal structure can therefore serve as a model for both Type I and Type II PCPs. Proteins 2014; 82:1210–1218. © 2013 Wiley Periodicals, Inc. 相似文献
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Ceri A. Fielding Rebecca Aicheler Richard J. Stanton Eddie C. Y. Wang Song Han Sepehr Seirafian James Davies Brian P. McSharry Michael P. Weekes P. Robin Antrobus Virginie Prod'homme Fabien P. Blanchet Daniel Sugrue Simone Cuff Dawn Roberts Andrew J. Davison Paul J. Lehner Gavin W. G. Wilkinson Peter Tomasec 《PLoS pathogens》2014,10(5)
NKG2D plays a major role in controlling immune responses through the regulation of natural killer (NK) cells, αβ and γδ T-cell function. This activating receptor recognizes eight distinct ligands (the MHC Class I polypeptide-related sequences (MIC) A andB, and UL16-binding proteins (ULBP)1–6) induced by cellular stress to promote recognition cells perturbed by malignant transformation or microbial infection. Studies into human cytomegalovirus (HCMV) have aided both the identification and characterization of NKG2D ligands (NKG2DLs). HCMV immediate early (IE) gene up regulates NKGDLs, and we now describe the differential activation of ULBP2 and MICA/B by IE1 and IE2 respectively. Despite activation by IE functions, HCMV effectively suppressed cell surface expression of NKGDLs through both the early and late phases of infection. The immune evasion functions UL16, UL142, and microRNA(miR)-UL112 are known to target NKG2DLs. While infection with a UL16 deletion mutant caused the expected increase in MICB and ULBP2 cell surface expression, deletion of UL142 did not have a similar impact on its target, MICA. We therefore performed a systematic screen of the viral genome to search of addition functions that targeted MICA. US18 and US20 were identified as novel NK cell evasion functions capable of acting independently to promote MICA degradation by lysosomal degradation. The most dramatic effect on MICA expression was achieved when US18 and US20 acted in concert. US18 and US20 are the first members of the US12 gene family to have been assigned a function. The US12 family has 10 members encoded sequentially through US12–US21; a genetic arrangement, which is suggestive of an ‘accordion’ expansion of an ancestral gene in response to a selective pressure. This expansion must have be an ancient event as the whole family is conserved across simian cytomegaloviruses from old world monkeys. The evolutionary benefit bestowed by the combinatorial effect of US18 and US20 on MICA may have contributed to sustaining the US12 gene family. 相似文献
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We have previously shown that whole cell preparations of Candida albicans are capable of inducing immunosuppressive B-cell activity both in vivo and in vitro. In an effort to characterize the components of the yeast which manifest this immunomodulatory activity, we have successfully generated a soluble extract with dithiothreitol which exerts immunosuppressive activity. This extract is capable of inducing antigen-nonspecific suppressor cells which inhibit the antibody response of normal cells in coculture. Both primary and secondary antibody responses are suppressed by these cells. Our results also show that the suppressor cell population is a member of the L3T4+ Ly-1+ Lyt-2- T-cell lineage. These results provide evidence that Candida extracts may possess clinically significant immunomodulatory activities. 相似文献
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Archibald Cuff 《BMJ (Clinical research ed.)》1908,1(2468):918-920