Mutations in the BRCT domain confer temperature sensitivity to BRCA1 in transcription activation.
BRCA1 is a tumor suppressor gene and germ line mutations account for the majority of familial cases of breast and ovarian cancer. There is mounting evidence that BRCA1 functions in DNA repair and transcriptional regulation. A major hurdle to dissect the role of BRCA1 is the lack of molecular reagents to carry out biochemical and genetic experiments. Therefore, we used random mutagenesis of the C-terminus of BRCA1 (aa 1560-1863) to generate temperature-sensitive (TS) mutants in transcription activation. We obtained 11 TS mutants in transcription that localized primarily to the hydrophobic core of the BRCT-N domain of BRCA1. One of the mutants, H1686Q, also displayed temperature-dependent transcription activation in human cells. These conditional mutants represent valuable tools to assess the role of BRCA1 in transcription activation.
Carvalho
Marcelo A
MA
Strang Cancer Prevention Center, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
Billack
Blase
B
Chan
Emily
E
Worley
Terri
T
Cayanan
Charmagne
C
Monteiro
Alvaro N A
AN
D000595:Amino Acid Sequence; D019313:BRCA1 Protein / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D001943:Breast Neoplasms / Q000209:etiology; D005260:Female; D006801:Humans; D008958:Models, Molecular; D009154:Mutation; D010051:Ovarian Neoplasms / Q000209:etiology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013696:Temperature; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D019313:BRCA1 Protein; D011993:Recombinant Fusion Proteins