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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
An WG, Kanekal M, Simon MC, Maltepe E, Blagosklonny MV and Neckers LM. . 1998 Nature 392: 405–408.
Andres JL, Fan S, Turkel GJ, Wang JA, Twu NF, Yuan RQ, Lamszus K, Goldberg ID and Rosen EM. . 1998 Oncogene 16: 2229–2241.
Andrews NC and Faller DV. . 1991 Nucleic Acids Res. 19: 2499.
Aniento F, Papavassiliou AG, Knecht E and Roche E. . 1996 FEBS Lett. 390: 47–52.
Aprelikova O, Kuthiala A, Bessho M, Ethier S and Liu ET. . 1996 Oncogene 13: 2487–2491.
Bishop DT and Kiemeney LA. . 1997 Semin. Cancer Biol. 8: 45–51.
Blackwood MA and Weber BL. . 1998 J. Clin. Oncol. 16: 1969–1977.
Blagosklonny MV. . 1997 Oncogene 15: 1889–1893.
Blagosklonny MV, Toretskey J and Neckers LM. . 1995 Oncogene 11: 933–939.
Brugarolas J and Jacks T. . 1997 Nature Med. 3: 721–722.
Carafoli E and Molinari M. . 1998 Biochem. Biophys. Res. Comm. 247: 193–203.
Chai YL, Cui J, Shao N, Shyam E, Reddy P and Rao VN. . 1999 Oncogene 18: 263–268.
Chapman MS and Verma IM. . 1996 Nature 382: 678–679.
Fan S, Wang J-A, Yuan R, Ma Y, Meng Q, Erdos MR, Pestell RG, Yuan F, Auborn KJ, Goldberg ID and Rosen EM. . 1999 Science 284: 1354–1356.
Fenteany G and Schreiber SL. . 1998 J. Biol. Chem. 273: 8545–8548.
Fornerod M, Ohno M, Yoshida M and Mattaj IW. . 1997 Cell 90: 1051–1060.
Freedman DA and Levine AJ. . 1998 Mol. Cell. Biol. 18: 7288–7293.
Fu Y-HF, Nishinaka T, Yokoyama K and Chiu R. . 1998 FEBS Lett. 421: 89–93.
Fukuda M, Asano S, Nakamura T, Adachi M, Yoshida M, Yanagida M and Nishida E. . 1997 Nature 390: 308–311.
Gonen H, Shkedy D, Barnoy S, Kosower NS and Ciechanover A. . 1997 FEBS Lett. 406: 17–22.
Grisham MB, Palombella VJ, Elliott PJ, Conner EM, Brand S, Wong HL, Pien C, Mazzola LM, Destree A, Parent L and Adams J. . 1999 Methods Enzymol. 300: 345–363.
Gudas JM, Li T, Nguyen H, Jensen D, Rauscher FJ and Cowan KH. . 1996 Cell Growth Differ. 7: 717–723.
Gudas JM, Nguyen H, Li T and Cowan KH. . 1995 Cancer Res. 55: 4561–4565.
Holaska JM and Paschal BM. . 1998 Proc. Natl. Acad. Sci. USA 95: 14739–14744.
Holt JT, Thompson ME, Szabo C, Tobinson-Benion C, Arteaga CL, King MC and Jensen RA. . 1996 Nature Genet. 12: 298–302.
Inoue S, Bar-Nun S, Roitelman J and Simoni RD. . 1991 J. Biol. Chem. 266: 13311–13317.
Jin Y, Xu XL, Yang MC, Wei F, Ayi TC, Bowcock AM and Baer R. . 1997 Proc. Natl. Acad. Sci. USA 94: 12075–12080.
Kastan MB, Onyekwere O, Sidransky D, Vogelstein B and Craig RW. . 1991 Cancer Res. 51: 6304–6311.
Kudo N, Wolff B, Sekimoto T, Schreiner EP, Yoneda Y, Yanagida M, Horinouchi S and Yoshida M. . 1998 Exp. Cell Res. 242: 540–547.
Kubbutat MH and Vousden KH. . 1997 Mol. Cell. Biol. 17: 460–468.
Lain S, Midgley C, Sparks A, Lane EB and Lane DP. . 1999 Exp. Cell Res. 248: 457–472.
Maki CG, Huibregtse JM and Howley PM. . 1996 Cancer Res. 56: 2649–2654.
Melillo G, Musso T, Sica A, Taylor LS, Cox GW and Varesio L. . 1995 J. Exp. Med. 182: 1683–1693.
Mercer WE and Baserga R. . 1985 Exp. Cell. Res. 160: 31–46.
Monteiro AN, August A and Hanafusa H. . 1996 Proc. Natl. Acad. Sci. USA 93: 13595–13599.
Nagaya T, Murata Y, Yamaguchi S, Nomura Y, Ohmori S, Fujieda M, Katunuma N, Yen PM, Chin WW and Seo H. . 1998 J. Biol. Chem. 273: 33166–33173.
Nelson WG and Kastan MB. . 1994 Mol. Cell. Biol. 14: 1815–1823.
Nishi K, Yoshida M, Fujiwara D, Nishikawa M, Horinouchi S and Beppu T. . 1994 J. Biol. Chem. 269: 6320–6324.
Nishinaka T, Fu Y-HF, Chen LI, Yokoyama K and Chiu R. . 1997 Biochim. Biophys. Acta 1351: 274–286.
Ouchi T, Monteiro AN, August A, Aaronson SA and Hanafusa H. . 1998 Proc. Natl. Acad. Sci. USA 95: 2302–2306.
Paoletti F, Mocali A and Tombaccini D. . 1997 J. Cell Physiol. 172: 63–68.
Pariat M, Carillo S, Molinari M, Salvat C, Debussche L, Bracco L, Milner J and Piechaczyk M. . 1997 Mol. Cell. Biol. 17: 2806–2815.
Reich NC and Levine AJ. . 1984 Nature 308: 199–201.
Rock KL, Gramm C, Rothstein L, Clark K, Stein R, Dick L, Hwang D and Goldberg AL. . 1994 Cell 78: 761–771.
Scheffner M, Werness BA, Huibregtse JM, Levine AJ and Howley PM. . 1990 Cell 63: 1129–1136.
Scheffner M. . 1998 Pharmacol. Ther. 78: 129–139.
Scully R, Chen J, Plug A, Xiao Y, Weaver D, Feunteun J, Ashley T and Livingston DM. . 1997 Cell 88: 265–275.
Somasundaram K, Zhang H, Zeng YX, Houvras Y, Peng Y, Zhang H, Wu GS, Licht JD, Weber BL and El-Deiry WS. . 1997 Nature 389: 187–190.
Stommel JM, Marchenko ND, Jimenez GS, Moll UM, Hope TJ and Wahl GM. . 1999 EMBO J. 18: 1660–1672.
Thompson ME, Jensen RA, Obermiller PS, Page DL and Holt JT. . 1995 Nat. Genet. 9: 444–450.
Wang KK, Nath R, Posner A, Raser KJ, Buroker-Kilgore M, Hajimohammadreza I, Probert AW, Marcoux FW, Ye Q, Takano E, Hatanaka M, Maki M, Caner H, Collins JL, Fergus A, Lee KS, Lunney EA, Hays SJ and Yuen P. . 1996 Proc. Natl. Acad. Sci. USA 93: 6687–6692.
Whitesell L, Sutphin P, An WG, Schulte T, Blagosklonny MV and Neckers L. . 1997 Oncogene 14: 2809–2816.
Zhang H, Somasundaram K, Peng Y, Tian H, Zhang H, Bi D, Weber BL and El-Deiry WS. . 1998 Oncogene 16: 1713–1721.
Zhang W, Lu Q, Xie ZJ and Mellgren RL. . 1997 Oncogene 14: 255–263.
Acknowledgements
We thank Peter Elliott for supplying the proteasome inhibitor PS341 and Minoru Yoshida for supplying leptomycin B.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Blagosklonny, M., An, W., Melillo, G. et al. Regulation of BRCA1 by protein degradation. Oncogene 18, 6460–6468 (1999). https://doi.org/10.1038/sj.onc.1203068
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1203068
Keywords
This article is cited by
-
Cisplatin-induced cell death increases the degradation of the MRE11-RAD50-NBS1 complex through the autophagy/lysosomal pathway
Cell Death & Differentiation (2023)
-
Regulating BRCA1 protein stability by cathepsin S-mediated ubiquitin degradation
Cell Death & Differentiation (2019)
-
NF-κB inhibition by dimethylaminoparthenolide radiosensitizes non-small-cell lung carcinoma by blocking DNA double-strand break repair
Cell Death Discovery (2018)
-
Zinc chelation induces rapid depletion of the X-linked inhibitor of apoptosis and sensitizes prostate cancer cells to TRAIL-mediated apoptosis
Cell Death & Differentiation (2008)
-
Role of BRCA1 in heat shock response
Oncogene (2003)