Document Detail


A role for DNA mismatch repair protein Msh2 in error-prone double-strand-break repair in mammalian chromosomes.
MedLine Citation:
PMID:  15781695     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We examined error-prone nonhomologous end joining (NHEJ) in Msh2-deficient and wild-type Chinese hamster ovary cell lines. A DNA substrate containing a thymidine kinase (tk) gene fused to a neomycin-resistance (neo) gene was stably integrated into cells. The fusion gene was rendered nonfunctional due to a 22-bp oligonucleotide insertion, which included the 18-bp I-SceI endonuclease recognition site, within the tk portion of the fusion gene. A double-strand break (DSB) was induced by transiently expressing the I-SceI endonuclease, and deletions or insertions that restored the tk-neo fusion gene's reading frame were recovered by selecting for G418-resistant colonies. Overall, neither the frequency of recovery of G418-resistant colonies nor the sizes of NHEJ-associated deletions were substantially different for the mutant vs. wild-type cell lines. However, we did observe greater usage of terminal microhomology among NHEJ events recovered from wild-type cells as compared to Msh2 mutants. Our results suggest that Msh2 influences error-prone NHEJ repair at the step of pairing of terminal DNA tails. We also report the recovery from both wild-type and Msh2-deficient cells of an unusual class of NHEJ events associated with multiple deletion intervals, and we discuss a possible mechanism for the generation of these "discontinuous deletions."
Authors:
Jason A Smith; Barbara Criscuolo Waldman; Alan S Waldman
Related Documents :
21094265 - Autophagy-related lipase fgatg15 of fusarium graminearum is important for lipid turnove...
16740945 - The orphan response regulator digr is required for synthesis of extracellular matrix fi...
15155625 - Role of flagellin and the two-component chea/chey system of listeria monocytogenes in h...
7665515 - Identification of a second cellulose synthase gene (acsaii) in acetobacter xylinum.
3858875 - Evolutionary amplification of a pseudogene.
15607945 - Engrailed genes control developmental fate of serotonergic and noradrenergic neurons in...
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.     Date:  2005-03-21
Journal Detail:
Title:  Genetics     Volume:  170     ISSN:  0016-6731     ISO Abbreviation:  Genetics     Publication Date:  2005 May 
Date Detail:
Created Date:  2005-05-25     Completed Date:  2006-06-06     Revised Date:  2013-06-09    
Medline Journal Info:
Nlm Unique ID:  0374636     Medline TA:  Genetics     Country:  United States    
Other Details:
Languages:  eng     Pagination:  355-63     Citation Subset:  IM    
Affiliation:
Department of Biological Sciences, University of South Carolina, Columbia, 29208, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Southern
CHO Cells
Cricetinae
Cricetulus
DNA Repair / physiology*
Polymerase Chain Reaction
Sequence Analysis, DNA
Grant Support
ID/Acronym/Agency:
GM47110/GM/NIGMS NIH HHS
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Elephants and human color-blind deuteranopes have identical sets of visual pigments.
Next Document:  Bayesian shrinkage estimation of quantitative trait loci parameters.