title: Regulation of BRCA1 by protein degradation.
abstract: BRCA1, a tumor suppressor protein implicated in hereditary forms of breast and ovarian cancer, is transcriptionally regulated in a proliferation-dependent manner. In this study, we demonstrate a substantial role for proteolysis in regulating the BRCA1 steady-state protein level in several cell lines. N-acetyl-leu-leu-norleucinal (ALLN), an inhibitor of the proteasome, calpain, and cathepsins, caused BRCA1 protein to accumulate in the nucleus of several human breast, prostate, and melanoma cell lines which express low or undetectable basal levels of BRCA1 protein, but not in cells with high basal expression of BRCA1. Protease inhibition did not increase BRCA1 synthesis, nor change its mRNA level, but it dramatically prolonged the protein's half-life. In contrast to ALLN, lactacystin and PS341, two specific proteasome inhibitors, as well as calpastatin peptide and PD150606, two selective calpain inhibitors, had no effect on BRCA1 stability, whereas ALLM, an effective calpain and cathepsin inhibitor but weak proteasome inhibitor, did stimulate accumulation of BRCA1. Moreover, three inhibitors of acidic cysteine proteases, chloroquine, ammonium chloride and bafilomycin, were as effective as ALLN. These results demonstrate that degradation by a cathepsin-like protease in fine balance with BRCA1 transcription is responsible for maintaining the low steady-state level of BRCA1 protein seen in many cancer cells.
authors.affiliation: Department of Therapeutics, National Cancer Institute, NIH, Bethesda, Maryland, MD 20892, USA.
authors.forename: M V
authors.initials: MV
authors.lastname: Blagosklonny
authors.forename: W G
authors.initials: WG
authors.lastname: An
authors.forename: G
authors.initials: G
authors.lastname: Melillo
authors.forename: P
authors.initials: P
authors.lastname: Nguyen
authors.forename: J B
authors.initials: JB
authors.lastname: Trepel
authors.forename: L M
authors.initials: LM
authors.lastname: Neckers
mesh_terms: D019313:BRCA1 Protein / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D002154:Calpain / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000378:metabolism; D003609:Dactinomycin / Q000037:antagonists & inhibitors; D015536:Down-Regulation / Q000187:drug effects; D010450:Endopeptidases / Q000378:metabolism; D005786:Gene Expression Regulation; D006207:Half-Life; D006801:Humans; D006868:Hydrolysis; D028884:Karyopherins; D007976:Leupeptins / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear; D014407:Tumor Cells, Cultured; D000097606:Exportin 1 Protein
chemical_list: D019313:BRCA1 Protein; D002154:Calpain; D002352:Carrier Proteins; D003546:Cysteine Endopeptidases; D003609:Dactinomycin; D010450:Endopeptidases; D028884:Karyopherins; D007976:Leupeptins; D009097:Multienzyme Complexes; D046988:Proteasome Endopeptidase Complex; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein; C063261:acetylleucyl-leucyl-norleucinal