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Molecular and Cellular Biology, July 1999, p. 4633-4642, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Interaction between the Product of the Breast Cancer
Susceptibility Gene BRCA2 and DSS1, a Protein
Functionally Conserved from Yeast to Mammals
Nicola J.
Marston,1
William J.
Richards,2
David
Hughes,2
David
Bertwistle,1
Christopher J.
Marshall,2 and
Alan
Ashworth1,*
Section of Gene Function and
Regulation1 and CRC Centre for Cell and
Molecular Biology,2 Institute of Cancer
Research, Chester Beatty Laboratories, London, United Kingdom SW3 6JB
Received 26 March 1999/Accepted 5 April 1999
Germ line mutations in the breast cancer susceptibility gene
BRCA2 predispose to early-onset breast cancer, but the
function of the nuclear protein encoded by the gene is ill defined.
Using the yeast two-hybrid system with fragments of human
BRCA2, we identified an interaction with the human
DSS1 (deleted in split hand/split foot) gene. Yeast and
mammalian two-hybrid assays showed that DSS1 can associate with BRCA2
in the region of amino acids 2472 to 2957 in the C terminus of the
protein. Using coimmunoprecipitation of epitope-tagged
BRCA2 and DSS1 cDNA constructs transiently
expressed in COS cells, we were able to demonstrate an
association. Furthermore, endogenous BRCA2 could be
coimmunoprecipitated with endogenous DSS1 in MCF7 cells, demonstrating
an in vivo association. Apparent orthologues of the mammalian
DSS1 gene were identified in the genome of the yeasts
Schizosaccharomyces pombe and Saccharomyces cerevisiae. Yeast strains in which these DSS1-like
genes were deleted showed a temperature-sensitive growth phenotype,
which was analyzed by flow cytometry. This provides evidence for a link between the BRCA2 tumor suppressor gene and a gene
required for completion of the cell cycle.
*
Corresponding author. Mailing address: Institute of
Cancer Research, Chester Beatty Laboratories, 237 Fulham Rd., London, United Kingdom SW3 6JB. Phone: (44)-171-352-8133. Fax:
(44)-171-352-3299. E-mail: alana{at}icr.ac.uk.
Molecular and Cellular Biology, July 1999, p. 4633-4642, Vol. 19, No. 7
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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