ACCA phosphopeptide recognition by the BRCT repeats of BRCA1.
The tumour suppressor gene BRCA1 encodes a 220 kDa protein that participates in multiple cellular processes. The BRCA1 protein contains a tandem of two BRCT repeats at its carboxy-terminal region. The majority of disease-associated BRCA1 mutations affect this region and provide to the BRCT repeats a central role in the BRCA1 tumour suppressor function. The BRCT repeats have been shown to mediate phospho-dependant protein-protein interactions. They recognize phosphorylated peptides using a recognition groove that spans both BRCT repeats. We previously identified an interaction between the tandem of BRCA1 BRCT repeats and ACCA, which was disrupted by germ line BRCA1 mutations that affect the BRCT repeats. We recently showed that BRCA1 modulates ACCA activity through its phospho-dependent binding to ACCA. To delineate the region of ACCA that is crucial for the regulation of its activity by BRCA1, we searched for potential phosphorylation sites in the ACCA sequence that might be recognized by the BRCA1 BRCT repeats. Using sequence analysis and structure modelling, we proposed the Ser1263 residue as the most favourable candidate among six residues, for recognition by the BRCA1 BRCT repeats. Using experimental approaches, such as GST pull-down assay with Bosc cells, we clearly showed that phosphorylation of only Ser1263 was essential for the interaction of ACCA with the BRCT repeats. We finally demonstrated by immunoprecipitation of ACCA in cells, that the whole BRCA1 protein interacts with ACCA when phosphorylated on Ser1263.
Ray
Hind
H
Laboratoire de Génétique Moléculaire, Signalisation et Cancer, CNRS UMR 5201, Faculté de Médecine Rockefeller, 8 Avenue Rockefeller, 69373 Lyon cedex 08, France.
Moreau
Karen
K
Dizin
Eva
E
Callebaut
Isabelle
I
Venezia
Nicole Dalla
ND
D000103:Acetyl-CoA Carboxylase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019313:BRCA1 Protein / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D006801:Humans; D047468:Immunoprecipitation; D008958:Models, Molecular; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011487:Protein Conformation; D020449:Repetitive Sequences, Amino Acid; D012694:Serine / Q000378:metabolism
D019313:BRCA1 Protein; D010750:Phosphoproteins; D012694:Serine; D000103:Acetyl-CoA Carboxylase