Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-XL

Wiese, Claudia; Pierce, Andrew J; Gauny, Stacey S; Jasin, Maria; Kronenberg, Amy

Title: Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-XL

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Abstract

Homology-directed repair (HDR) of DNA double-strand breaks (DSBs) is a well-established mechanism that contributes to the maintenance of genomic stability in rodent cells, and it has been assumed that HDR is of similar importance in the repair of DSBs in human cells. However, in addition to promoting genomic stability, some outcomes of homologous recombination can be deleterious, suggesting that factors exist to regulate HDR. We previously demonstrated that overexpression of BCL-2 or BCL-xL enhanced the frequency of x-ray-induced mutations involving the TK1 locus, including loss of heterozygosity (LOH) events presumed to arise by mitotic recombination. The present study was designed to test whether HDR is a prominent DSB repair pathway in human cells, and to directly determine whether ectopic expression of BCL-xL affects HDR. We used the B-lymphoblastoid cell line TK6, which expresses wild-type TP53 and resembles normal lymphocytes in undergoing apoptosis following! genotoxic stress. U sing isogenic derivatives of TK6 cells (TK6-neo, TK6-bcl-xL), we find that a DSB in an integrated HDR reporter stimulates gene conversion 40-50-fold in TK6-neo cells, demonstrating that a DSB can be efficiently repaired by gene conversion in human cells. Significantly, DSB-induced gene conversion events are 3- to 4-fold more frequent in BCL-xL overexpressing cells.more »« less

Authors:: Wiese, Claudia; Pierce, Andrew J; Gauny, Stacey S; Jasin, Maria; Kronenberg, Amy

Publication Date:: Tue Sep 25 00:00:00 EDT 2001

Research Org.:: Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

Sponsoring Org.:: USDOE Director, Office of Science; National Aeronautics and Space Administration (US)

OSTI Identifier:: 795338

Report Number(s):: LBNL-49067
Journal ID: ISSN 0008-5472; CNREA8; R&D Project: 865915; TRN: US200212%%63

DOE Contract Number:: AC03-76SF00098

Resource Type:: Journal Article

Journal Name:: Cancer Research

Additional Journal Information:: Journal Volume: 62; Journal Issue: 5; Other Information: Journal Publication Date: 2002 Mar. 1; PBD: 25 Sep 2001; Journal ID: ISSN 0008-5472

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; ANIMAL CELLS; APOPTOSIS; BIOLOGICAL PATHWAYS; DNA; GENES; LYMPHOCYTES; MUTATIONS; GENE RECOMBINATION; BIOLOGICAL REPAIR; RODENTS; HUMAN POPULATIONS; DOUBLE-STRAND BREAK REPAIR HOMOLOGOUS RECOMBINATION BCL-XL APOPTOSIS HUMAN CELLS

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                    Wiese, Claudia, Pierce, Andrew J, Gauny, Stacey S, Jasin, Maria, and Kronenberg, Amy. Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-XL.  United States: N. p., 2001. 
        Web.

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                    Wiese, Claudia, Pierce, Andrew J, Gauny, Stacey S, Jasin, Maria, & Kronenberg, Amy. Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-XL.  United States.

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                    Wiese, Claudia, Pierce, Andrew J, Gauny, Stacey S, Jasin, Maria, and Kronenberg, Amy. 2001.  
        "Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-XL".  United States.  https://www.osti.gov/servlets/purl/795338.

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@article{osti_795338,

  title        = {Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-XL},

  author       = {Wiese, Claudia and Pierce, Andrew J and Gauny, Stacey S and Jasin, Maria and Kronenberg, Amy},

  abstractNote = {Homology-directed repair (HDR) of DNA double-strand breaks (DSBs) is a well-established mechanism that contributes to the maintenance of genomic stability in rodent cells, and it has been assumed that HDR is of similar importance in the repair of DSBs in human cells. However, in addition to promoting genomic stability, some outcomes of homologous recombination can be deleterious, suggesting that factors exist to regulate HDR. We previously demonstrated that overexpression of BCL-2 or BCL-xL enhanced the frequency of x-ray-induced mutations involving the TK1 locus, including loss of heterozygosity (LOH) events presumed to arise by mitotic recombination. The present study was designed to test whether HDR is a prominent DSB repair pathway in human cells, and to directly determine whether ectopic expression of BCL-xL affects HDR. We used the B-lymphoblastoid cell line TK6, which expresses wild-type TP53 and resembles normal lymphocytes in undergoing apoptosis following! genotoxic stress. U sing isogenic derivatives of TK6 cells (TK6-neo, TK6-bcl-xL), we find that a DSB in an integrated HDR reporter stimulates gene conversion 40-50-fold in TK6-neo cells, demonstrating that a DSB can be efficiently repaired by gene conversion in human cells. Significantly, DSB-induced gene conversion events are 3- to 4-fold more frequent in BCL-xL overexpressing cells. The results demonstrate that HDR plays an important role in maintaining genomic integrity in human cells and that ectopic expression of BCL-xL enhances HDR of DSBs. To our knowledge, this is the first study to highlight a function for BCL-xL in modulating DSB repair in human cells.},

  doi          = {},

  url          = {https://www.osti.gov/biblio/795338},
  journal      = {Cancer Research},

  issn         = {0008-5472},

  number       = 5,

  volume       = 62,

  place        = {United States},

  year         = {Tue Sep 25 00:00:00 EDT 2001},

  month        = {Tue Sep 25 00:00:00 EDT 2001}

}

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