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(Received for publication, May 10, 1996, and in revised form, September 20, 1996)
From The breast cancer susceptibility gene
BRCA1 encodes an 1863-amino acid protein that acts as a
tumor suppressor. The biochemical function of BRCA1 is unknown, and
there are conflicting results describing its subcellular location. We
have identified a 220-kDa protein, which is reactive with three
antibodies raised against the amino- and carboxyl-terminal regions of
BRCA1. Immunoflourescence staining with an antibody to the carboxyl
terminus of BRCA1 localized the protein to the nucleus of breast,
ovarian, and cervical carcinoma-derived cell lines. A similar result
was observed by biochemical subcellular fractionation that indicated
that the 220-kDa protein was localized primarily to the nucleus of cell
lines established from breast carcinomas. In addition to the 220-kDa
protein, one antibody, C-20, also recognized a 180-kDa protein in
MDA-MB-468 total cell lysates that was not detected by the other two
antibodies. Several observations suggest the 180-kDa protein is the
epidermal growth factor (EGF) receptor: (i) C-20 reacted avidly with a
180-kDa protein immunoprecipitated by an antibody to the EGF receptor;
(ii) an EGF receptor antibody detected a 180-kDa protein
immunoprecipitated by C-20; (iii) the affinity purified EGF receptor
was both immunoprecipitated and detected on immunoblots by the C-20
antibody but not another BRCA1 antibody; (iv) similar phosphopeptide
maps were generated from the EGF receptor and the 180-kDa protein
immunoprecipitated by C-20, and this peptide map was distinct from the
220-kDa phosphoprotein; and (v) the C-20 immunizing peptide bears
sequence identity to the EGF receptor. These results indicate that
BRCA1 is a 220-kDa nuclear protein and that the 180-kDa protein
reported previously may be unrelated to BRCA1.
Volume 271, Number 45,
Issue of November 8, 1996
pp. 28630-28635
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
,
,
,
and
Cancer Research, Lilly Research Laboratories,
Eli Lilly and Co., Lilly Corporate Center, Indianapolis, Indiana 46285
and ¶ Onyx Pharmaceuticals, Inc., Richmond, California 94806
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