Roles of Caenorhabditis elegans WRN Helicase in DNA Damage Responses, and a Comparison with Its Mammalian Homolog: A Mini-Review

Gerontology ◽  
2015 ◽  
Vol 62 (3) ◽  
pp. 296-303 ◽  
Author(s):  
Jin-Sun Ryu ◽  
Hyeon-Sook Koo
Keyword(s):  
Dna Damage ◽  
Caenorhabditis Elegans ◽  
Werner Syndrome ◽  
Model Organisms ◽  
Current Status ◽  
Helicase Domain ◽  
Replication Forks ◽  
Dna Breaks ◽  
Double Strand Dna Breaks ◽  
Double Strand Dna

Werner syndrome protein (WRN) is unusual among RecQ family DNA helicases in having an additional exonuclease activity. WRN is involved in the repair of double-strand DNA breaks via the homologous recombination and nonhomologous end joining pathways, and also in the base excision repair pathway. In addition, the protein promotes the recovery of stalled replication forks. The helicase activity is thought to unwind DNA duplexes, thereby moving replication forks or Holliday junctions. The targets of the exonuclease could be the nascent DNA strands at a replication fork or the ends of double-strand DNA breaks. However, it is not clear which enzyme activities are essential for repairing different types of DNA damage. Model organisms such as mice, flies, and worms deficient in WRN homologs have been investigated to understand the physiological results of defects in WRN activity. Premature aging, the most remarkable characteristic of Werner syndrome, is also seen in the mutant mice and worms, and hypersensitivity to DNA damage has been observed in WRN mutants of all three model organisms, pointing to conservation of the functions of WRN. In the nematode Caenorhabditis elegans, the WRN homolog contains a helicase domain but no exonuclease domain, so that this animal is very useful for studying the in vivo functions of the helicase without interference from the activity of the exonuclease. Here, we review the current status of investigations of C. elegans WRN-1 and discuss its functional differences from the mammalian homologs.

scholarly journals STRIDE—a fluorescence method for direct, specific in situ detection of individual single- or double-strand DNA breaks in fixed cells

2019 ◽  
Vol 48 (3) ◽  
pp. e14-e14 ◽  
Author(s):  
Magdalena M Kordon ◽  
Mirosław Zarębski ◽  
Kamil Solarczyk ◽  
Hanhui Ma ◽  
Thoru Pederson ◽  
...  
Keyword(s):  
Dna Damage ◽  
Single Cells ◽  
Dna Lesions ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Dna Breaks ◽  
Damage Site ◽  
Double Strand Dna Breaks ◽  
In Situ Detection ◽  
Double Strand Dna

Abstract We here describe a technique termed STRIDE (SensiTive Recognition of Individual DNA Ends), which enables highly sensitive, specific, direct in situ detection of single- or double-strand DNA breaks (sSTRIDE or dSTRIDE), in nuclei of single cells, using fluorescence microscopy. The sensitivity of STRIDE was tested using a specially developed CRISPR/Cas9 DNA damage induction system, capable of inducing small clusters or individual single- or double-strand breaks. STRIDE exhibits significantly higher sensitivity and specificity of detection of DNA breaks than the commonly used terminal deoxynucleotidyl transferase dUTP nick-end labeling assay or methods based on monitoring of recruitment of repair proteins or histone modifications at the damage site (e.g. γH2AX). Even individual genome site-specific DNA double-strand cuts induced by CRISPR/Cas9, as well as individual single-strand DNA scissions induced by the nickase version of Cas9, can be detected by STRIDE and precisely localized within the cell nucleus. We further show that STRIDE can detect low-level spontaneous DNA damage, including age-related DNA lesions, DNA breaks induced by several agents (bleomycin, doxorubicin, topotecan, hydrogen peroxide, UV, photosensitized reactions) and fragmentation of DNA in human spermatozoa. The STRIDE methods are potentially useful in studies of mechanisms of DNA damage induction and repair in cell lines and primary cultures, including cells with impaired repair mechanisms.

scholarly journals Direct, sensitive and specific detection of individual single- or double-strand DNA breaks by fluorescence microscopy

2019 ◽  
Author(s):  
Magdalena Kordon ◽  
Mirosław Zarębski ◽  
Kamil Solarczyk ◽  
Hanhui Ma ◽  
Thoru Pederson ◽  
...  
Keyword(s):  
Dna Damage ◽  
Fluorescence Microscopy ◽  
Single Cells ◽  
Primary Cultures ◽  
Dna Lesions ◽  
Dna Breaks ◽  
Damage Site ◽  
Double Strand Dna Breaks ◽  
Age Related ◽  
Double Strand Dna

ABSTRACTWe here describe a technique termed STRIDE (SensiTive Recognition of Individual DNA Ends), which enables highly sensitive, specific, direct in situ detection of single- or double-strand DNA breaks (sSTRIDE or dSTRIDE), in nuclei of single cells, using fluorescence microscopy. Sensitivity of STRIDE was tested using specially developed CRISPR/Cas9 DNA damage induction system, capable of inducing small clusters or individual single- or double-strand breaks. STRIDE exhibits significantly higher sensitivity and specificity of detection of DNA breaks than the commonly used TUNEL assay or methods based on monitoring of recruitment of repair proteins or histone modifications at the damage site (e.g. γH2AX). Even individual genome site-specific DNA double-strand cuts induced by CRISPR/Cas9, as well as individual single-strand DNA scissions induced by the nickase version of Cas9, can be detected by STRIDE and precisely localized within the cell nucleus. We further show that STRIDE can detect low-level spontaneous DNA damage, including age-related DNA lesions, DNA breaks induced by several agents (bleomycin, doxorubicin, topotecan, hydrogen peroxide, UV, photosensitized reactions), and fragmentation of DNA in human spermatozoa. STRIDE methods are potentially useful in studies of mechanisms of DNA damage induction and repair in cell lines and primary cultures, including cells with impaired repair mechanisms.

scholarly journals Single and Double Strand Sperm DNA Damage: Different Reproductive Effects on Male Fertility

Genes ◽  
2019 ◽  
Vol 10 (2) ◽  
pp. 105 ◽  
Author(s):  
Jordi Ribas-Maynou ◽  
Jordi Benet
Keyword(s):  
Dna Damage ◽  
Male Fertility ◽  
Sperm Chromatin ◽  
Dna Breaks ◽  
Sperm Dna Damage ◽  
Reproductive Effects ◽  
Double Strand Dna Breaks ◽  
Sperm Dna ◽  
Break Points ◽  
Double Strand Dna

Reproductive diseases have become a growing worldwide problem and male factor plays an important role in the reproductive diagnosis, prognosis and design of assisted reproductive treatments. Sperm cell holds the mission of carrying the paternal genetic complement to the oocyte in order to contribute to an euploid zygote with proper DNA integrity. Sperm DNA fragmentation had been used for decades as a male fertility test, however, its usefulness have arisen multiple debates, especially around Intracytoplasmic Sperm Injection (ICSI) treatments. In the recent years, it has been described that different types of sperm DNA breaks (single and double strand DNA breaks) cause different clinical reproductive effects. On one hand, single-strand DNA breaks are present extensively as a multiple break points in all regions of the genome, are related to oxidative stress and cause a lack of clinical pregnancy or an increase of the conception time. On the other hand, double-strand DNA breaks are mainly localized and attached to the sperm nuclear matrix as a very few break points, are possibly related to a lack of DNA repair in meiosis and cause a higher risk of miscarriage, low embryo quality and higher risk of implantation failure in ICSI cycles. The present work also reviews different studies that may contribute in the understanding of sperm chromatin as well as treatments to prevent sperm DNA damage.

scholarly journals p53 Prevents the Accumulation of Double-Strand DNA Breaks at Stalled-Replication Forks Induced by UV in Human Cells

Cell Cycle ◽  
2004 ◽  
Vol 3 (12) ◽  
pp. 1543-1557 ◽  
Author(s):  
Shoshana Squires ◽  
Julia A. Coates ◽  
Michal Goldberg ◽  
Lorraine H. Toji ◽  
Stephen P. Jackson ◽  
...  
Keyword(s):  
Human Cells ◽  
Replication Forks ◽  
Dna Breaks ◽  
Double Strand Dna Breaks ◽  
Stalled Replication Forks ◽  
Double Strand Dna

scholarly journals Fine-Structure Mapping of Meiosis-Specific Double-Strand DNA Breaks at a Recombination Hotspot Associated With an Insertion of Telomeric Sequences Upstream of the HIS4 Locus in Yeast

Genetics ◽  
1996 ◽  
Vol 143 (3) ◽  
pp. 1115-1125 ◽  
Author(s):  
Fei Xu ◽  
Thomas D Petes
Keyword(s):  
Meiotic Recombination ◽  
Hydroxyl Groups ◽  
Sequence Specificity ◽  
Structure Mapping ◽  
Dna Breaks ◽  
Exonuclease Iii ◽  
Recombination Hotspot ◽  
Telomeric Sequences ◽  
Double Strand Dna Breaks ◽  
Double Strand Dna

Abstract Meiotic recombination in Saccharomyces cerevisiae is initiated by double-strand DNA breaks (DSBs). Using two approaches, we mapped the position of DSBs associated with a recombination hotspot created by insertion of telomeric sequences into the region upstream of HIS4. We found that the breaks have no obvious sequence specificity and localize to a region of ~50 bp adjacent to the telomeric insertion. By mapping the breaks and by studies of the exonuclease III sensitivity of the broken ends, we conclude that most of the broken DNA molecules have blunt ends with 3′-hydroxyl groups.

Expression of double strand DNA breaks repair genes in pterygium

2010 ◽  
Vol 32 (1) ◽  
pp. 39-47 ◽  
Author(s):  
Anna Łękawa–Ilczuk ◽  
Halina Antosz ◽  
Beata Rymgayłło–Jankowska ◽  
Tomasz Żarnowski
Keyword(s):  
Dna Breaks ◽  
Double Strand Dna Breaks ◽  
Repair Genes ◽  
Double Strand Dna

Inside Back Cover: Double-Strand DNA Breaks Induced by Paracyclophane Gold(I) Complexes (Chem. Eur. J. 26/2017)

2017 ◽  
Vol 23 (26) ◽  
pp. 6459-6459
Author(s):  
Sebastian Bestgen ◽  
Carmen Seidl ◽  
Thomas Wiesner ◽  
Andreas Zimmer ◽  
Martina Falk ◽  
...  
Keyword(s):  
Dna Breaks ◽  
Back Cover ◽  
Double Strand Dna Breaks ◽  
Double Strand Dna
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