Skip to main content
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1996 Dec;87(12):1204–1209. doi: 10.1111/j.1349-7006.1996.tb03134.x

Failure to Detect Mutations in the Retinoblastoma Protein‐binding Domain of the Transcription Factor E2F‐1 in Human Cancers

Takao Nakamura 1, Yoshiaki Monden 1,, Kazuko Kawashima 2, Tsuguo Naruke 3, Susumu Nishimura 1
PMCID: PMC5921028  PMID: 9045954

Abstract

The functions of the transcription factor E2F‐1 are regulated by the RB protein through the RB‐binding domain of E2F‐1 and this factor is considered to be an important molecule that functions downstream of the RB protein. In order to determine whether E2F‐1 that cannot bind to RB might be associated with various human cancers, we searched for mutations in the RB‐binding domain of E2F‐1 using samples of DNA from various clinical specimens obtained from 406 cancer patients (with lung, pancreatic, stomach, colon, esophageal, and hepatic cancers) by analysis of polymerase chain reaction‐mediated single‐strand conformational polymorphism. No mutations or deletions were detected in genes for E2F‐1 from any of the tumor tissues examined. These results suggest that a mutation or deletion in E2F‐1 that might affect binding of the RB protein is not involved in human cancers.

Keywords: Retinoblastoma protein, E2F‐1, Mutation, Carcinogenesis, PCR‐SSCP

Full Text

The Full Text of this article is available as a PDF (334.3 KB).

REFERENCES

  • 1. ) Goodrich , D. W. , Wang , N. P. , Qian , Y.‐W. , Lee , E. Y.‐H. P. and Lee , W.‐H.The retinoblastoma gene product regulates progression through the G1 phase of the cell cycle . Cell , 67 , 293 – 302 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Weinberg , R. A.The retinoblastoma protein and cell cycle control . Cell , 81 , 323 – 330 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Kaelin , W. G. , Jr. , Pallas , D. C. , DeCaprio , J. A. , Kaye , F. J. and Livingston , D. M.Identification of cellular proteins that can interact specifically with the T/E1A‐ binding region of the retinoblastoma gene product . Cell , 64 , 521 – 532 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 4. ) Huang , S. , Wang , N.‐P. , Tseng , B. Y. , Lee , W.‐H. and Lee , E. Y.‐ H. P.Two distinct and frequently mutated regions of retinoblastoma protein are required for binding to SV40 T antigen . EMBO J. , 9 , 1815 – 1822 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5. ) Kaelin , W. G. , Jr. , Ewen , M. E. and Livingston , D. M.Definition of the minimal simian virus 40 large T antigen‐ and adenovirus E1A‐binding domain in the retinoblastoma gene product . Mol. Cell. Biol. , 10 , 3761 – 3769 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6. ) Bookstein , R. , Shew , J.‐Y. , Chen , P.‐L. , Scully , P. and Lee , W.‐H.Suppression of tumorigenicity of human prostate carcinoma cells by replacing a mutated RB gene . Science , 247 , 712 – 715 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Hu , Q. , Dyson , N. and Harlow , E.The regions of the retinoblastoma protein needed for binding to adenovirus E1A or SV40 large T antigen are common sites for mutations . EMBO J. , 9 , 1147 – 1155 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8. ) Yokota , J. , Akiyama , T. , Fung , Y.‐K. T. , Benedict , W. F. , Namba , Y. , Hanaoka , M. , Wada , M. , Terasaki , T. , Shimosato , Y. , Sugimura , T. and Terada , M.Altered expression of the retinoblastoma (RB) gene in small‐cell carcinoma of the lung . Oncogene , 3 , 471 – 475 ( 1988. ). [PubMed] [Google Scholar]
  • 9. ) DeCaprio , J. A. , Ludlow , J. W. , Figge , J. , Shew , J.‐Y. , Huang , C.‐M. , Lee , W.‐H. , Marsilio , E. , Paucha , E. and Livingston , D. M.SV40 large tumor antigen forms a specific complex with the product of the retinoblastoma susceptibility gene . Cell , 54 , 275 – 283 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) Chellappan , S. P. , Hiebert , S. , Mudryj , M. , Horowitz , J. M. and Nevins , J. R.The E2F transcription factor is a cellular target for the RB protein . Cell , 65 , 1053 – 1061 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 11. ) Cobrinik , D. , Whyte , P. , Peeper , D. S. , Jacks , T. and Weinberg , R. A.Cell cycle‐specific association of E2F with the p130 E1A‐binding protein . Genes Dev. , 7 , 2392 – 2404 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Blake , M. C. and Azizkhan , J. C.Transcription factor E2F is required for efficient expression of the hamster dihydrofolate reductase gene in vitro and in vivo . Mol. Cell. Biol. , 9 , 4994 – 5002 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13. ) Pearson , B. E. , Nasheuer , H.‐P. and Wang , T. S.‐F.Human DNA polymerase α gene: sequences controlling expression in cycling and serum‐stimulated cells . Mol. Cell. Biol. , 11 , 2081 – 2095 ( 1991. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. ) Dalton , S.Cell cycle regulation of the human cdc2 gene . EMBO J. , 11 , 1797 – 1804 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. ) Shan , B. , Chang , C.‐Y. , Jones , D. and Lee , W.‐H.The transcription factor E2F‐1 mediates the autoregulation of RB gene expression . Mol. Cell. Biol. , 14 , 299 – 309 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. ) Kim , Y. K. and Lee , A. S.Identification of a 70‐base‐pair cell cycle regulatory unit within the promoter of the human thymidine kinase gene and its interaction with cellular factors . Mol. Cell. Biol. , 11 , 2296 – 2302 ( 1991. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. ) Hiebert , S. W. , Lipp , M. and Nevins , J. R.E1A‐dependent trans‐activation of the human MYC promoter is mediated by the E2F factor . Proc. Natl. Acad. Sci. USA , 86 , 3594 – 3598 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. ) Lam , E. W.‐F. and Watson , R. J.An E2F‐binding site mediates cell‐cycle regulated repression of mouse B‐myb transcription . EMBO J. , 12 , 2705 – 2713 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. ) Nevins , J. R.E2F: a link between the RB tumor suppressor protein and viral oncoproteins . Science , 258 , 424 – 429 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Helin , K. , Harlow , E. and Fattaey , A.Inhibition of E2F‐1 transactivation by direct binding of the retinoblastoma protein . Mol. Cell. Biol. , 13 , 6501 – 6508 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. ) Hiebert , S. W. , Chellappan , S. P. , Horowitz , J. M. and Nevins , J. R.The interaction of RB with E2F coincides with an inhibition of the transcriptional activity of E2F . Genes Dev. , 6 , 177 – 185 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Weintraub , S. J. , Prater , C. A. and Dean , D. C.Retinoblastoma protein switches the E2F site from positive to negative element . Nature , 358 , 259 – 261 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Qin , X.‐Q. , Livingston , D. M. , Ewen , M. , Sellers , W. R. , Arany , Z. and Kaelin , W. G. , Jr.The transcription factor E2F‐1 is a downstream target of RB action . Mol. Cell. Biol. , 15 , 742 – 755 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. ) Lukas , J. , Petersen , B. O. , Holm , K. , Bartek , J. and Helin , K.Deregulated expression of E2F family members induces S‐phase entry and overcomes p16INK4A‐mediated growth suppression . Mol. Cell. Biol. , 16 , 1047 – 1057 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. ) Orita , M. , Suzuki , Y. , Sekiya , T. and Hayashi , K.Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction . Genomics , 5 , 874 – 879 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Blin , N. and Stafford , D. W.A general method for isolation of high molecular weight DNA from eukaryotes . Nucleic Acids Res. , 3 , 2303 – 2308 ( 1976. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27. ) Kovesdi , I. , Reichel , R. and Nevins , J. R.Identification of a cellular transcription factor involved in E1A trans‐activation . Cell , 45 , 219 – 228 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Xu , G. , Livingston , D. M. and Krek , W.Multiple members of the E2F transcription factor family are the products of oncogenes . Proc. Natl. Acad. Sci. USA , 92 , 1357 – 1361 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29. ) Johnson , D. G. , Cress , W. D. , Jakoi , L. and Nevins , J. R.Oncogenic capacity of the E2F1 gene . Proc. Natl. Acad. Sci. USA , 91 , 12823 – 12827 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30. ) Berger , R. , Bloomfield , C. D. and Sutherland , G. R.Human gene mapping 8: report of the committee on chromosome rearrangements in neoplasia and on fragile sites . Cytogenet. Cell Genet. , 40 , 490 – 535 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Yamasaki , L. , Jacks , T. , Bronson , R. , Goillot , E. , Harlow , E. and Dyson , N. J.Tumor induction and tissue atrophy in mice lacking E2F‐1 . Cell , 85 , 537 – 548 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Field , S. J. , Tsai , F.‐Y. , Kuo , F. , Zubiaga , A. M. , Kaelin , W. G. , Jr. , Livingston , D. M. , Orkin , S. H. and Greenberg , M. E.E2F‐1 functions in mice to promote apoptosis and suppress proliferation . Cell , 85 , 549 – 561 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Shiseki , M. , Kohno , T. , Nishikawa , R. , Sameshima , Y. , Mizoguchi , H. and Yokota , J.Frequent allelic losses on chromosomes 2q, 18q, and 22q in advanced non‐small cell lung carcinoma . Cancer Res. , 54 , 5643 – 5648 ( 1994. ). [PubMed] [Google Scholar]
  • 34. ) Waber , P. G. , Chen , J. and Nisen , P. D.Infrequency of ras, p53, WT1, or RB gene alterations in Wilms tumors . Cancer , 72 , 3732 – 3738 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 35. ) Yokota , J. and Sugimura , T.Multiple steps in carcinogenesis involving alterations of multiple tumor suppressor genes . FASEB J. , 7 , 920 – 925 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 36. ) Hesketh , R.“The Oncogene Handbook,” pp. 378 – 413 , 525 – 553 ( 1994. ). Academic Press; , London . [Google Scholar]
  • 37. ) Bishop , J. M.Molecular themes in oncogenesis . Cell , 64 , 235 – 248 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 38. ) Bos , J. L.ras oncogenes in human cancer . Cancer Res. , 49 , 4682 – 4689 ( 1989. ). [PubMed] [Google Scholar]
  • 39. ) Utida , T. , Watanabe , T. , Kinoshita , T. , Murate , T. , Saito , H. and Hotta , T.Mutational analysis of the CDKN2 (MTS1/p16INK4A) gene in primary B‐cell lymphomas . Blood , 86 , 2724 – 2731 ( 1995. ). [PubMed] [Google Scholar]
  • 40. ) Shimizu , T. , Miwa , W. , Nakamori , S. , Ishikawa , O. , Konishi , Y. and Sekiya , T.Absence of a mutation of the p21/WAF1 gene in human lung and pancreatic cancers . Jpn. J. Cancer Res. , 87 , 275 – 278 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41. ) Miwa , W. , Yasuda , J. , Yashima , K. , Makino , R. and Sekiya , T.Absence of activating mutations of the RAF1 protooncogene in human lung cancer . Biol. Chem. Hoppe-Seyler , 375 , 705 – 709 ( 1994. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES