Reduced X-ray resistance and homologous recombination frequencies in a RAD54(-/-) mutant of the chicken DT40 cell line

被引：236

作者：

Bezzubova, O ^{[1
]}

Silbergleit, A ^{[1
]}

YamaguchiIwai, Y ^{[1
]}

Takeda, S ^{[1
]}

Buerstedde, JM ^{[1
]}

机构：

[1] KYOTO UNIV, SCH MED, BAYER CHAIR DEPT MOL IMMUNOL, KYOTO 606, JAPAN

来源：

CELL | 1997年 / 89卷 / 02期

关键词：

D O I：

10.1016/S0092-8674(00)80198-1

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

rad54 mutants of the yeast Saccharomyces cerevisiae are extremely X-ray sensitive and have decreased mitotic recombination frequencies because of a defect in double-strand break repair. A RAD54 homolog was disrupted in the chicken B cell line DT40, which undergoes immunoglobulin gene conversion and exhibits unusually high ratios of targeted to random integration after DNA transfection. Homozygous RAD54(-/-) mutant clones were highly X-ray sensitive compared to wildtype cells. The rate of immunoglobulin gene conversion was 6- to 8-fold reduced, and the frequency of targeted integration was at least two orders of magnitude decreased in the mutant clones. Reexpression of the RAD54 cDNA restored radiation resistance and targeted integration activity. The reported phenotype provides the first genetic evidence of a link between double-strand break repair and homologous recombination in vertebrate cells.

引用

页码：185 / 193

页数：9

共 49 条

[1] IDENTIFICATION OF A MOUSE HOMOLOG OF THE SACCHAROMYCES-CEREVISIAE RECOMBINATION AND REPAIR GENE, RAD52 [J].

BENDIXEN, C ;

SUNJEVARIC, I ;

BAUCHWITZ, R ;

ROTHSTEIN, R .

GENOMICS, 1994, 23 (01) :300-303

[2] PURIFICATION AND CHARACTERIZATION OF THE HUMAN RAD51 PROTEIN, AN ANALOG OF ESCHERICHIA-COLI RECA [J].

BENSON, FE ;

STASIAK, A ;

WEST, SC .

EMBO JOURNAL, 1994, 13 (23) :5764-5771

[3] A CHICKEN RAD51 HOMOLOG IS EXPRESSED AT HIGH-LEVELS IN LYMPHOID AND REPRODUCTIVE-ORGANS [J].

BEZZUBOVA, O ;

SHINOHARA, A ;

MUELLER, RG ;

OGAWA, H ;

BUERSTEDDE, JM .

NUCLEIC ACIDS RESEARCH, 1993, 21 (07) :1577-1580

[4] GENE CONVERSION IN THE CHICKEN IMMUNOGLOBULIN LOCUS - A PARADIGM OF HOMOLOGOUS RECOMBINATION IN HIGHER EUKARYOTES [J].

BEZZUBOVA, OY ;

BUERSTEDDE, JM .

EXPERIENTIA, 1994, 50 (03) :270-276

[5] IDENTIFICATION OF A CHICKEN RAD52 HOMOLOG SUGGESTS CONSERVATION OF THE RAD52 RECOMBINATION PATHWAY THROUGHOUT THE EVOLUTION OF HIGHER EUKARYOTES [J].

BEZZUBOVA, OY ;

SCHMIDT, H ;

OSTERMANN, K ;

HEYER, WD ;

BUERSTEDDE, JM .

NUCLEIC ACIDS RESEARCH, 1993, 21 (25) :5945-5949

[6] DEFECTIVE DNA-DEPENDENT PROTEIN-KINASE ACTIVITY IS LINKED TO V(D)J RECOMBINATION AND DNA-REPAIR DEFECTS ASSOCIATED WITH THE MURINE SCID MUTATION [J].

BLUNT, T ;

FINNIE, NJ ;

TACCIOLI, GE ;

SMITH, GCM ;

DEMENGEOT, J ;

GOTTLIEB, TM ;

MIZUTA, R ;

VARGHESE, AJ ;

ALT, FW ;

JEGGO, PA ;

JACKSON, SP .

CELL, 1995, 80 (05) :813-823

[7] REPAIR OF DOUBLE-STRAND BREAKS IN A TEMPERATURE CONDITIONAL RADIATION-SENSITIVE MUTANT OF SACCHAROMYCES CEREVISIAE [J].

BUDD, M ;

MORTIMER, RK .

MUTATION RESEARCH, 1982, 103 (01) :19-24

[8] INCREASED RATIO OF TARGETED TO RANDOM INTEGRATION AFTER TRANSFECTION OF CHICKEN B-CELL LINES [J].

BUERSTEDDE, JM ;

TAKEDA, S .

CELL, 1991, 67 (01) :179-188

[9] LIGHT CHAIN GENE CONVERSION CONTINUES AT HIGH-RATE IN AN ALV-INDUCED CELL-LINE [J].

BUERSTEDDE, JM ;

REYNAUD, CA ;

HUMPHRIES, EH ;

OLSON, W ;

EWERT, DL ;

WEILL, JC .

EMBO JOURNAL, 1990, 9 (03) :921-927

[10] ISOLATION AND CHARACTERIZATION OF YEAST DNA-REPAIR GENES .2. ISOLATION OF PLASMIDS THAT COMPLEMENT THE MUTATIONS RAD50-1, RAD51-1, RAD54-3, AND RAD55-3 [J].

CALDERON, IL ;

CONTOPOULOU, CR ;

MORTIMER, RK .

CURRENT GENETICS, 1983, 7 (02) :93-100

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