scholarly journals Mutations in the WRN Gene in Mice Accelerate Mortality in a p53-Null Background

2000 ◽  
Vol 20 (9) ◽  
pp. 3286-3291 ◽  
Author(s):  
David B. Lombard ◽  
Caroline Beard ◽  
Brad Johnson ◽  
Robert A. Marciniak ◽  
Jessie Dausman ◽  
...  
Keyword(s):  
Mortality Rate ◽  
Life Span ◽  
Human Disease ◽  
Dna Helicase ◽  
Premature Aging ◽  
Mutant Mice ◽  
Helicase Domain ◽  
C Terminus ◽  
Accelerated Senescence

ABSTRACT Werner's syndrome (WS) is a human disease with manifestations resembling premature aging. The gene defective in WS, WRN, encodes a DNA helicase. Here, we describe the generation of mice bearing a mutation that eliminates expression of the C terminus of the helicase domain of the WRN protein. Mutant mice are born at the expected Mendelian frequency and do not show any overt histological signs of accelerated senescence. These mice are capable of living beyond 2 years of age. Cells from these animals do not show elevated susceptibility to the genotoxins camptothecin or 4-NQO. However, mutant fibroblasts senesce approximately one passage earlier than controls. Importantly,WRN−/− ;p53−/− mice show an increased mortality rate relative toWRN+/− ;p53−/− animals. We consider possible models for the synergy betweenp53 and WRN mutations for the determination of life span.

scholarly journals The Short Life Span of Saccharomyces cerevisiae sgs1 and srs2 Mutants Is a Composite of Normal Aging Processes and Mitotic Arrest Due to Defective Recombination

Genetics ◽  
2001 ◽  
Vol 157 (4) ◽  
pp. 1531-1542 ◽  
Author(s):  
Mitch McVey ◽  
Matt Kaeberlein ◽  
Heidi A Tissenbaum ◽  
Leonard Guarente
Keyword(s):  
Cell Cycle ◽  
Life Span ◽  
Mitotic Cell ◽  
Dna Helicase ◽  
Premature Aging ◽  
Growth Defect ◽  
Short Life Span ◽  
Single Strand Annealing ◽  
Late Generation ◽  
Strand Annealing

Abstract Evidence from many organisms indicates that the conserved RecQ helicases function in the maintenance of genomic stability. Mutation of SGS1 and WRN, which encode RecQ homologues in budding yeast and humans, respectively, results in phenotypes characteristic of premature aging. Mutation of SRS2, another DNA helicase, causes synthetic slow growth in an sgs1 background. In this work, we demonstrate that srs2 mutants have a shortened life span similar to sgs1 mutants. Further dissection of the sgs1 and srs2 survival curves reveals two distinct phenomena. A majority of sgs1 and srs2 cells stops dividing stochastically as large-budded cells. This mitotic cell cycle arrest is age independent and requires the RAD9-dependent DNA damage checkpoint. Late-generation sgs1 and srs2 cells senesce due to apparent premature aging, most likely involving the accumulation of extrachromosomal rDNA circles. Double sgs1 srs2 mutants are viable but have a high stochastic rate of terminal G2/M arrest. This arrest can be suppressed by mutations in RAD51, RAD52, and RAD57, suggesting that the cell cycle defect in sgs1 srs2 mutants results from inappropriate homologous recombination. Finally, mutation of RAD1 or RAD50 exacerbates the growth defect of sgs1 srs2 cells, indicating that sgs1 srs2 mutants may utilize single-strand annealing as an alternative repair pathway.

scholarly journals Human WRN is an intrinsic inhibitor of progerin, abnormal splicing product of lamin A

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
So-mi Kang ◽  
Min-Ho Yoon ◽  
Su-Jin Lee ◽  
Jinsook Ahn ◽  
Sang Ah Yi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Life Span ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Werner Syndrome ◽  
Genetic Disorder ◽  
Dna Helicase ◽  
Premature Aging ◽  
Lamin A ◽  
Short Life Span

AbstractWerner syndrome (WRN) is a rare progressive genetic disorder, caused by functional defects in WRN protein and RecQ4L DNA helicase. Acceleration of the aging process is initiated at puberty and the expected life span is approximately the late 50 s. However, a Wrn-deficient mouse model does not show premature aging phenotypes or a short life span, implying that aging processes differ greatly between humans and mice. Gene expression analysis of WRN cells reveals very similar results to gene expression analysis of Hutchinson Gilford progeria syndrome (HGPS) cells, suggesting that these human progeroid syndromes share a common pathological mechanism. Here we show that WRN cells also express progerin, an abnormal variant of the lamin A protein. In addition, we reveal that duplicated sequences of human WRN (hWRN) from exon 9 to exon 10, which differ from the sequence of mouse WRN (mWRN), are a natural inhibitor of progerin. Overexpression of hWRN reduced progerin expression and aging features in HGPS cells. Furthermore, the elimination of progerin by siRNA or a progerin-inhibitor (SLC-D011 also called progerinin) can ameliorate senescence phenotypes in WRN fibroblasts and cardiomyocytes, derived from WRN-iPSCs. These results suggest that progerin, which easily accumulates under WRN-deficient conditions, can lead to premature aging in WRN and that this effect can be prevented by SLC-D011.

scholarly journals A Conserved Regulatory Module at the C Terminus of the Papillomavirus E1 Helicase Domain Controls E1 Helicase Assembly

2014 ◽  
Vol 89 (2) ◽  
pp. 1129-1142 ◽  
Author(s):  
Stephen Schuck ◽  
Arne Stenlund
Keyword(s):  
Inhibitory Effect ◽  
Dna Helicase ◽  
Regulatory Sequence ◽  
Helicase Domain ◽  
E1 Protein ◽  
C Terminus ◽  
Regulatory Module ◽  
Origin Of Dna Replication ◽  
Coding Capacity ◽  
Regulatory Challenges

ABSTRACTViruses frequently combine multiple activities into one polypeptide to conserve coding capacity. This strategy creates regulatory challenges to ascertain that the combined activities are compatible and do not interfere with each other. The papillomavirus E1 protein, as many other helicases, has the intrinsic ability to form hexamers and double hexamers (DH) that serve as the replicative DNA helicase. However, E1 also has the more unusual ability to generate local melting by forming a double trimer (DT) complex that can untwist the double-stranded origin of DNA replication (ori) DNA in preparation for DH formation. Here we describe a switching mechanism that allows the papillomavirus E1 protein to form these two different kinds of oligomers and to transition between them. We show that a conserved regulatory module attached to the E1 helicase domain blocks hexamer and DH formation and promotes DT formation. In the presence of the appropriate trigger, the inhibitory effect of the regulatory module is relieved and the transition to DH formation can occur.IMPORTANCEThis study provides a mechanistic understanding into how a multifunctional viral polypeptide can provide different, seemingly incompatible activities. A conserved regulatory sequence module attached to the AAA+helicase domain in the papillomavirus E1 protein allows the formation of different oligomers with different biochemical activities.

scholarly journals The Main Role of Srs2 in DNA Repair Depends on Its Helicase Activity, Rather than on Its Interactions with PCNA or Rad51

mBio ◽  
2018 ◽  
Vol 9 (4) ◽  
Author(s):  
Alex Bronstein ◽  
Lihi Gershon ◽  
Gilad Grinberg ◽  
Elisa Alonso-Perez ◽  
Martin Kupiec
Keyword(s):  
Dna Damage ◽  
Dna Replication ◽  
Genome Stability ◽  
Dna Helicase ◽  
Main Role ◽  
Helicase Domain ◽  
Helicase Activity ◽  
C Terminus ◽  
Srs2 Helicase

ABSTRACTHomologous recombination (HR) is a mechanism that repairs a variety of DNA lesions. Under certain circumstances, however, HR can generate intermediates that can interfere with other cellular processes such as DNA transcription or replication. Cells have therefore developed pathways that abolish undesirable HR intermediates. TheSaccharomyces cerevisiaeyeast Srs2 helicase has a major role in one of these pathways. Srs2 also works during DNA replication and interacts with the clamp PCNA. The relative importance of Srs2’s helicase activity, Rad51 removal function, and PCNA interaction in genome stability remains unclear. We created a newSRS2allele [srs2(1-850)] that lacks the whole C terminus, containing the interaction site for Rad51 and PCNA and interactions with many other proteins. Thus, the new allele encodes an Srs2 protein bearing only the activity of the DNA helicase. We find that the interactions of Srs2 with Rad51 and PCNA are dispensable for the main role of Srs2 in the repair of DNA damage in vegetative cells and for proper completion of meiosis. On the other hand, it has been shown that in cells impaired for the DNA damage tolerance (DDT) pathways, Srs2 generates toxic intermediates that lead to DNA damage sensitivity; we show that this negative Srs2 activity requires the C terminus of Srs2. Dissection of the genetic interactions of thesrs2(1-850) allele suggest a role for Srs2’s helicase activity in sister chromatid cohesion. Our results also indicate that Srs2’s function becomes more central in diploid cells.IMPORTANCEHomologous recombination (HR) is a key mechanism that repairs damaged DNA. However, this process has to be tightly regulated; failure to regulate it can lead to genome instability. The Srs2 helicase is considered a regulator of HR; it was shown to be able to evict the recombinase Rad51 from DNA. Cells lacking Srs2 exhibit sensitivity to DNA-damaging agents, and in some cases, they display defects in DNA replication. The relative roles of the helicase and Rad51 removal activities of Srs2 in genome stability remain unclear. To address this question, we created a new Srs2 mutant which has only the DNA helicase domain. Our study shows that only the DNA helicase domain is needed to deal with DNA damage and assist in DNA replication during vegetative growth and in meiosis. Thus, our findings shift the view on the role of Srs2 in the maintenance of genome integrity.

scholarly journals Structure determination of Murine Norovirus NS6 proteases with C-terminal extensions designed to probe protease-substrate interactions

2014 ◽  
Author(s):  
Humberto Fernandes ◽  
Eoin N Leen ◽  
Hamlet Cromwell Jnr ◽  
Marc-Philipp Pfeil ◽  
Stephen Curry
Keyword(s):  
Structure Determination ◽  
Crystal Form ◽  
Molecular Replacement ◽  
Murine Norovirus ◽  
Substrate Interactions ◽  
C Terminus ◽  
Peptide Binding Site ◽  
Specific Protease ◽  
Protease Substrate

Noroviruses are positive-sense single-stranded RNA viruses. They encode an NS6 protease that cleaves a viral polyprotein at specific sites to produce mature viral proteins. In an earlier study we obtained crystals of murine norovirus (MNV) NS6 protease in which crystal contacts were mediated by specific insertion of the C-terminus of one protein (which contains residues P5-P1 of the NS6-7 cleavage junction) into the peptide binding site of an adjacent molecule, forming an adventitious protease-product complex. We sought to reproduce this crystal form to investigate protease-substrate complexes by extending the C-terminus of NS6 construct to include residues on the C-terminal (P´) side of the cleavage junction. We report the crystallization and crystal structure determination of inactive mutants of murine norovirus NS6 protease with C-terminal extensions of one, two and four residues from the N-terminus of the adjacent NS7 protein (NS6 1´, NS6 2´, NS6 4´). We also determined the structure of a chimeric extended NS6 protease in which the P4 P4′ sequence of the NS6-7 cleavage site was replaced with the corresponding sequence from the NS2-3 cleavage junction (NS6 4´ 2|3). The constructs NS6 1′ and NS6 2′ yielded crystals that diffracted anisotropically. We found that, although the uncorrected data could be phased by molecular replacement, refinement of the structures stalled unless the data were ellipsoidally truncated and corrected with anisotropic B-factors. These corrections significantly improved phasing by molecular replacement and subsequent refinement. The refined structures of all four extended NS6 proteases are very similar in structure to the mature MNV NS6 — and in one case reveal additional details of a surface loop. Although the packing arrangement observed showed some similarities to those observed in the adventitious protease-product crystals reported previously, in no case were specific protease-substrate interactions observed.

The amino acid sequence of yeast phosphoglycerate mutase

1982 ◽  
Vol 215 (1198) ◽  
pp. 19-44 ◽  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Proteolytic Enzymes ◽  
Phosphoglycerate Mutase ◽  
Crystallographic Structure ◽  
X Ray ◽  
C Terminus ◽  
Detailed Interpretation ◽  
High Degree

The complete amino acid sequence of yeast phosphoglycerate mutase comprising 241 residues has been determined. The sequence was deduced from the two cyanogen bromide fragments, and from the peptides derived from these fragments after digestion by a number of proteolytic enzymes. Determination of this sequence now allows a detailed interpretation of the existing high-resolution X-ray crystallographic structure. A comparison of the sequence reported here with the sequences of peptides from phosphoglycerate mutases from other species, and with the sequence of erythrocyte diphosphoglycerate mutase, indicates that these enzymes have a high degree of structural homology. Autolysis of phosphoglycerate mutase by yeast extracts leads to the complete loss of mutase activity, and the formation of electrophoretically distinguishable forms (R. Sasaki, E. Sugimoto & H. Chiba, Archs Biochem. Biophys. 115, 53-61 (1966)). It is apparent from the amino acid sequence that these changes are due to the loss of an 8─12 residue peptide from the C-terminus.

Periventricular Intraparenchymal Echodensities in the Premature Newborn: Critical Determinant of Neurologic Outcome

PEDIATRICS ◽  
1986 ◽  
Vol 78 (6) ◽  
pp. 995-1006
Author(s):  
Franco Guzzetta ◽  
Gary D. Shackelford ◽  
Sara Volpe ◽  
Jeffrey M. Perlman ◽  
Joseph J. Volpe
Keyword(s):  
Cognitive Function ◽  
Mortality Rate ◽  
Cognitive Outcome ◽  
Motor Deficits ◽  
Critical Determinant ◽  
Acute Period ◽  
Hemorrhagic Necrosis ◽  
Neurologic Disability ◽  
Basic Characteristics

Controversy exists concerning the degree of importance of periventricular intraparenchymal echodensities (IPE) observed on neonatal ultrasound scans in the determination of subsequent neurologic disability in premature infants. In this report, IPE was studied in 75 infants weighing less than 2,000 g at birth to determine the basic characteristics of the lesion, the likely pathogenesis, the outcome, and the aspects of the ultrasonographic appearance in the acute period of neonatal illness that are important for prediction of outcome. IPE was defined as any periventricular echodensity greater than 1 cm in at least one dimension. IPE was strikingly associated with large areas of intraventricular hemorrhage (IVH) (81% of cases). IPE was distinctly asymmetric. Thus, the lesion was either exclusively unilateral (67%) or bilateral with marked predominance on one side. The associated IVH was asymmetric in approximately 80% of cases, and in all 50 cases of large asymmetric IVH, IPE occurred on the same side as the larger amount of intraventricular blood. Moreover, more than 50% of such cases of IPE associated with large asymmetric IVH were progressive. Neuropathologic correlation showed that IPE represented hemorrhagic necrosis of periventricular tissue. Concerning pathogenesis, these data raise the possibility that large asymmetric IVH is related etiologically to IPE. Outcome varied with the severity of the IPE. Thus, the mortality rate among the 38 infants with extensive IPE was 79%. Of the survivors with extensive IPE, all had subsequent major motor deficits and all but one exhibited cognitive function less than 80% of normal. Among the 37 infants with localized IPE, the mortality rate was 38%. Of the survivors, although 79% had major motor deficits, 43% had cognitive function greater than 80% of normal. Thus, the findings demonstrate that with extensive IPE there is little or no chance for survival with normal neurologic and cognitive outcome, but with localized IPE, although major motor deficits are common, an appreciable proportion of infants have cognitive function in the normal range. Careful, quantitative assessment of the ultrasonographic features of IPE in the acute period of illness in the premature infant is of major value in estimating outcome.

Determination of impact of Azadirachta indica L. leaves on adults and eggs of Callosobruchus maculatus Fabricius, the largest predator of stored cowpea (Vigna unguiculata Walp) by an applicable method by farmers

2015 ◽  
Vol 4 (5) ◽  
pp. 215-221
Author(s):  
Ablaye Faye ◽  
Malick Sarr ◽  
Abdoulaye Samb ◽  
Cheikh Thiaw ◽  
Mbacké Sembène
Keyword(s):  
Mortality Rate ◽  
Callosobruchus Maculatus ◽  
Maximum Effect ◽  
Maximal Effect ◽  
Cowpea Weevil ◽  
Lethal Effects ◽  
Neem Leaves ◽  
Leaf Powder ◽  
Higher Doses

  The effect of neem leaves has been tested in the laboratory on eggs and adults of cowpea weevil (C. maculatus). Different formulations of this plant were applied to these forms of C. maculatus Fab. Grinding fresh contact sheets induced significant lethal effects from 96.12% to 100% on eggs; whereas 100% of mortality was recorded at the end of eight days of applica-tion to three adults with higher doses. Fumigation on turn proved less effec-tive than contact on eggs. It induced a maximal effect of 95.73% mortality with the larger dose (D4: 0.02912g/cm3). On adults, we recorded highest mortality (100%) from the 7th day of the show with the highest dose. The aqueous extract of neem leaf powder was less effective than all other formu-lations on the eggs as well as adults of this insect; with a higher mortality rate (74.99%) observed on the eggs with the application of the concentration C2. On adults we recorded a maximum effect (100% mortality) from the 13th day of the application with the highest concentration (C1). These mortalities would be related to the support of several active molecules contained in neem as established in literature.

scholarly journals Methods for Quantification of the Decline Phenomenon and Determination of the Vulnerability Degree for the Oak Stands in Northwestern Transylvania, Romania

2017 ◽  
Vol 45 (2) ◽  
pp. 623-631
Author(s):  
Vasile ŞIMONCA ◽  
Ioan OROIAN ◽  
Dănuț CHIRA ◽  
Ioan TĂUT
Keyword(s):  
Mortality Rate ◽  
Incipient Stage ◽  
Management Measures ◽  
Slow Development ◽  
Synthetic Index ◽  
Quercus Species ◽  
Fast Development ◽  
Annual Mortality ◽  
Experimental Plots

The decline phenomenon of stands can be defined as the totality of effects that damage the normal vegetation state of trees and stands. There are two types of decline, the acute one, with fast development, and the chronical one, with slow development in space and time. The accurate capture of the phenomenon has direct implications for the forest management measures. A first step in this direction is the analysis of the development type and of its dynamics. Forests which have Quercus species in composition are affected more by this phenomenon. There were identified oak stands presenting decline phenomenon in 9 Forest Districts in Northwestern Transylvania and 22 experimental plots were analyzed. These plots were located in middle aged stands that were affected in different ways. The study was performed during 2013-2015 and focused on crown level assessments of trees, taking into consideration the shape and size, defoliations, percentage of dead or missing slender branches, number of dead thick branches, presence of epicormic branches or dormant buds on the stem, presence of pathogens or pests. These elements were used for calculating some technical indicators of the health and vegetation state: degree of dieback, damage and devitalization, synthetic index damage, annual mortality rate. In 7 out of 22 stands, the overcoming of the incipient stage and beginning of active dieback stage was observed. The strongest correlation occurred between the annual mortality rate and the synthetic index damage.

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