Open AccessHuman BRCA1 inhibits growth in yeast: Potential use in diagnostic testing
Author(s) -
Jeffrey S. Humphrey,
Ali Salim,
Michael R. Erdos,
Francis S. Collins,
Lawrence C. Brody,
Richard D. Klausner
Publication year - 1997
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.94.11.5820
Subject(s) - missense mutation , frameshift mutation , biology , genetics , saccharomyces cerevisiae , nonsense mutation , fusion protein , mutation , yeast , germline , gene , recombinant dna
Germline-inactivating mutations ofBRCA1 result in a hereditary predisposition to breast and ovarian cancer. Truncating mutations of BRCA1 predispose to cancer and can be ascertained by protein truncation testing or sequencing. However, cancer-predisposing missense mutations ofBRCA1 are difficult to distinguish from polymorphisms by genetic testing methods currently used. Here we show that expression ofBRCA1 orBRCA1 fused to aGAL4 activation domain inSaccharomyces cerevesiae inhibits growth, resulting in small colonies easily distinguishable from vector-transformed controls. The growth inhibitory effect can be localized to sequences encoding the recently described BRCA1 C-terminal domains. Growth suppression by a BRCA1 fusion protein is not influenced by introduction of neutral polymorphisms but is diminished or abolished by frameshift, nonsense, or disease-associated missense mutations located in the C-terminal 305 amino acids of BRCA1. These observations may permit the functional significance of manyBRCA1 sequence changes to be assessed in yeast. Additionally, the correlation of growth suppression with wild-type forms of BRCA1 suggests that the assay may be capable of detecting functionally conserved interactions between the evolutionarily conserved BRCA1 C-terminal domains and cellular elements found in both human and yeast cells.