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. 1997 Jan;17(1):444–452. doi: 10.1128/mcb.17.1.444

Localization of BRCA1 and a splice variant identifies the nuclear localization signal.

S Thakur 1, H B Zhang 1, Y Peng 1, H Le 1, B Carroll 1, T Ward 1, J Yao 1, L M Farid 1, F J Couch 1, R B Wilson 1, B L Weber 1
PMCID: PMC231769  PMID: 8972225

Abstract

Inherited mutations in BRCA1 confer susceptibility to breast and ovarian neoplasms. However, the function of BRCA1 and the role of BRCA1 in noninherited cancer remain unknown. Characterization of alternately spliced forms of BRCA1 may identify functional regions; thus, we constructed expression vectors of BRCA1 and a splice variant lacking exon 11, designated BRCA1 delta 672-4095. Immunofluorescence studies indicate nuclear localization of BRCA1 but cytoplasmic localization of BRCA1 delta 672-4095. Two putative nuclear localization signals (designated NLS1 and NLS2) were identified in exon 11; immunofluorescence studies indicate that only NLS1 is required for nuclear localization. RNA analysis indicates the expression of multiple, tissue-specific forms of BRCA1 RNAs; protein analysis with multiple antibodies suggests that at least three BRCA1 isoforms are expressed, including those lacking exon 11. The results suggest that BRCA1 is a nuclear protein and raise the possibility that splicing is one form of regulation of BRCA1 function by alteration of the subcellular localization of expressed proteins.

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Selected References

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  1. Abel K. J., Xu J., Yin G. Y., Lyons R. H., Meisler M. H., Weber B. L. Mouse Brca1: localization sequence analysis and identification of evolutionarily conserved domains. Hum Mol Genet. 1995 Dec;4(12):2265–2273. doi: 10.1093/hmg/4.12.2265. [DOI] [PubMed] [Google Scholar]
  2. Beckmann H., Su L. K., Kadesch T. TFE3: a helix-loop-helix protein that activates transcription through the immunoglobulin enhancer muE3 motif. Genes Dev. 1990 Feb;4(2):167–179. doi: 10.1101/gad.4.2.167. [DOI] [PubMed] [Google Scholar]
  3. Bennett L. M., Haugen-Strano A., Cochran C., Brownlee H. A., Fiedorek F. T., Jr, Wiseman R. W. Isolation of the mouse homologue of BRCA1 and genetic mapping to mouse chromosome 11. Genomics. 1995 Oct 10;29(3):576–581. doi: 10.1006/geno.1995.9963. [DOI] [PubMed] [Google Scholar]
  4. Boulikas T. Nuclear localization signals (NLS). Crit Rev Eukaryot Gene Expr. 1993;3(3):193–227. [PubMed] [Google Scholar]
  5. Castilla L. H., Couch F. J., Erdos M. R., Hoskins K. F., Calzone K., Garber J. E., Boyd J., Lubin M. B., Deshano M. L., Brody L. C. Mutations in the BRCA1 gene in families with early-onset breast and ovarian cancer. Nat Genet. 1994 Dec;8(4):387–391. doi: 10.1038/ng1294-387. [DOI] [PubMed] [Google Scholar]
  6. Chen Y., Chen C. F., Riley D. J., Allred D. C., Chen P. L., Von Hoff D., Osborne C. K., Lee W. H. Aberrant subcellular localization of BRCA1 in breast cancer. Science. 1995 Nov 3;270(5237):789–791. doi: 10.1126/science.270.5237.789. [DOI] [PubMed] [Google Scholar]
  7. Chen Y., Chen P. L., Riley D. J., Lee W. H., Allred D. C., Osborne C. K. Response: Location of BRCA1 in Human Breast and Ovarian Cancer Cells. Science. 1996 Apr 5;272(5258):125–126. doi: 10.1126/science.272.5258.125. [DOI] [PubMed] [Google Scholar]
  8. Couch F. J., Weber B. L. Mutations and polymorphisms in the familial early-onset breast cancer (BRCA1) gene. Breast Cancer Information Core. Hum Mutat. 1996;8(1):8–18. doi: 10.1002/humu.1380080102. [DOI] [PubMed] [Google Scholar]
  9. Easton D. F., Bishop D. T., Ford D., Crockford G. P. Genetic linkage analysis in familial breast and ovarian cancer: results from 214 families. The Breast Cancer Linkage Consortium. Am J Hum Genet. 1993 Apr;52(4):678–701. [PMC free article] [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  11. Freemont P. S. The RING finger. A novel protein sequence motif related to the zinc finger. Ann N Y Acad Sci. 1993 Jun 11;684:174–192. doi: 10.1111/j.1749-6632.1993.tb32280.x. [DOI] [PubMed] [Google Scholar]
  12. Futreal P. A., Liu Q., Shattuck-Eidens D., Cochran C., Harshman K., Tavtigian S., Bennett L. M., Haugen-Strano A., Swensen J., Miki Y. BRCA1 mutations in primary breast and ovarian carcinomas. Science. 1994 Oct 7;266(5182):120–122. doi: 10.1126/science.7939630. [DOI] [PubMed] [Google Scholar]
  13. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  14. Gudas J. M., Nguyen H., Li T., Cowan K. H. Hormone-dependent regulation of BRCA1 in human breast cancer cells. Cancer Res. 1995 Oct 15;55(20):4561–4565. [PubMed] [Google Scholar]
  15. Hannink M., Temin H. M. Transactivation of gene expression by nuclear and cytoplasmic rel proteins. Mol Cell Biol. 1989 Oct;9(10):4323–4336. doi: 10.1128/mcb.9.10.4323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Holt J. T., Thompson M. E., Szabo C., Robinson-Benion C., Arteaga C. L., King M. C., Jensen R. A. Growth retardation and tumour inhibition by BRCA1. Nat Genet. 1996 Mar;12(3):298–302. doi: 10.1038/ng0396-298. [DOI] [PubMed] [Google Scholar]
  17. Jensen R. A., Thompson M. E., Jetton T. L., Szabo C. I., van der Meer R., Helou B., Tronick S. R., Page D. L., King M. C., Holt J. T. BRCA1 is secreted and exhibits properties of a granin. Nat Genet. 1996 Mar;12(3):303–308. doi: 10.1038/ng0396-303. [DOI] [PubMed] [Google Scholar]
  18. Kalderon D., Richardson W. D., Markham A. F., Smith A. E. Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature. 1984 Sep 6;311(5981):33–38. doi: 10.1038/311033a0. [DOI] [PubMed] [Google Scholar]
  19. Kiledjian M., Su L. K., Kadesch T. Identification and characterization of two functional domains within the murine heavy-chain enhancer. Mol Cell Biol. 1988 Jan;8(1):145–152. doi: 10.1128/mcb.8.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marquis S. T., Rajan J. V., Wynshaw-Boris A., Xu J., Yin G. Y., Abel K. J., Weber B. L., Chodosh L. A. The developmental pattern of Brca1 expression implies a role in differentiation of the breast and other tissues. Nat Genet. 1995 Sep;11(1):17–26. doi: 10.1038/ng0995-17. [DOI] [PubMed] [Google Scholar]
  21. Miki Y., Swensen J., Shattuck-Eidens D., Futreal P. A., Harshman K., Tavtigian S., Liu Q., Cochran C., Bennett L. M., Ding W. A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. Science. 1994 Oct 7;266(5182):66–71. doi: 10.1126/science.7545954. [DOI] [PubMed] [Google Scholar]
  22. Morin N., Delsert C., Klessig D. F. Nuclear localization of the adenovirus DNA-binding protein: requirement for two signals and complementation during viral infection. Mol Cell Biol. 1989 Oct;9(10):4372–4380. doi: 10.1128/mcb.9.10.4372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rao V. N., Shao N., Ahmad M., Reddy E. S. Antisense RNA to the putative tumor suppressor gene BRCA1 transforms mouse fibroblasts. Oncogene. 1996 Feb 1;12(3):523–528. [PubMed] [Google Scholar]
  24. Rexach M., Blobel G. Protein import into nuclei: association and dissociation reactions involving transport substrate, transport factors, and nucleoporins. Cell. 1995 Dec 1;83(5):683–692. doi: 10.1016/0092-8674(95)90181-7. [DOI] [PubMed] [Google Scholar]
  25. Scully R., Ganesan S., Brown M., De Caprio J. A., Cannistra S. A., Feunteun J., Schnitt S., Livingston D. M. Location of BRCA1 in human breast and ovarian cancer cells. Science. 1996 Apr 5;272(5258):123–126. doi: 10.1126/science.272.5258.123. [DOI] [PubMed] [Google Scholar]
  26. Sharan S. K., Wims M., Bradley A. Murine Brca1: sequence and significance for human missense mutations. Hum Mol Genet. 1995 Dec;4(12):2275–2278. doi: 10.1093/hmg/4.12.2275. [DOI] [PubMed] [Google Scholar]
  27. Slavicek J. M., Jones N. C., Richter J. D. A karyophilic signal sequence in adenovirus type 5 E1A is functional in Xenopus oocytes but not in somatic cells. J Virol. 1989 Sep;63(9):4047–4050. doi: 10.1128/jvi.63.9.4047-4050.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thompson M. E., Jensen R. A., Obermiller P. S., Page D. L., Holt J. T. Decreased expression of BRCA1 accelerates growth and is often present during sporadic breast cancer progression. Nat Genet. 1995 Apr;9(4):444–450. doi: 10.1038/ng0495-444. [DOI] [PubMed] [Google Scholar]
  29. Van Etten R. A., Jackson P., Baltimore D. The mouse type IV c-abl gene product is a nuclear protein, and activation of transforming ability is associated with cytoplasmic localization. Cell. 1989 Aug 25;58(4):669–678. doi: 10.1016/0092-8674(89)90102-5. [DOI] [PubMed] [Google Scholar]

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