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    
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."
Jason A Smith; Barbara Criscuolo Waldman; Alan S Waldman
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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    
Department of Biological Sciences, University of South Carolina, Columbia, 29208, USA.
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MeSH Terms
Blotting, Southern
CHO Cells
DNA Repair / physiology*
Polymerase Chain Reaction
Sequence Analysis, DNA
Grant Support

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