Deletion of Brca2 exon 27 causes hypersensitivity to DNA crosslinks, chromosomal instability, and reduced life span in mice.
The Brca2 tumor-suppressor gene contributes to genomic stability, at least in part by a role in homologous recombinational repair. BRCA2 protein is presumed to function in homologous recombination through interactions with RAD51. Both exons 11 and 27 of Brca2 code for domains that interact with RAD51; exon 11 encodes eight BRC motifs, whereas exon 27 encodes a single, distinct interaction domain. Deletion of all RAD51-interacting domains causes embryonic lethality in mice. A less severe phenotype is seen with BRAC2 truncations that preserve some, but not all, of the BRC motifs. These mice can survive beyond weaning, but are runted and infertile, and die very young from cancer. Cells from such mice show hypersensitivity to some genotoxic agents and chromosomal instability. Here, we have analyzed mice and cells with a deletion of only the RAD51-interacting region encoded by exon 27. Mice homozygous for this mutation (called brca2(lex1)) have a shorter life span than that of control littermates, possibly because of early onsets of cancer and sepsis. No other phenotype was observed in these animals; therefore, the brca2(lex1) mutation is less severe than truncations that delete some BRC motifs. However, at the cellular level, the brca2(lex1) mutation causes reduced viability, hypersensitivity to the DNA interstrand crosslinking agent mitomycin C, and gross chromosomal instability, much like more severe truncations. Thus, the extreme carboxy-terminal region encoded by exon 27 is important for BRCA2 function, probably because it is required for a fully functional interaction between BRCA2 and RAD51.
Donoho
Greg
G
Lexicon Genetics, Inc., The Woodlands, Texas, USA.
Brenneman
Mark A
MA
Cui
Tracy X
TX
Donoviel
Dorit
D
Vogel
Hannes
H
Goodwin
Edwin H
EH
Chen
David J
DJ
Hasty
Paul
P
D000818:Animals; D024682:BRCA2 Protein / Q000235:genetics* / Q000378:metabolism; D001947:Breeding; D002460:Cell Line; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000235:genetics; D002873:Chromosome Fragility / Q000235:genetics*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D018736:DNA Adducts / Q000235:genetics*; D004249:DNA Damage / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D005091:Exons / Q000235:genetics*; D005720:Gamma Rays; D024522:Genes, BRCA2; D008136:Longevity / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D016685:Mitomycin / Q000494:pharmacology; D051135:Rad51 Recombinase; D017384:Sequence Deletion
D024682:BRCA2 Protein; D003432:Cross-Linking Reagents; D018736:DNA Adducts; D004268:DNA-Binding Proteins; D016685:Mitomycin; D051135:Rad51 Recombinase; C495154:Rad51 protein, mouse