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. 2000 Oct;91(10):987–993. doi: 10.1111/j.1349-7006.2000.tb00875.x

Overexpression of DA41 in v‐Ha‐ras‐3Y1 Cells Causes Growth Suppression

Toshinori Ozaki 1,✉, Yohko Nakamura 1, Eiji Hanaoka 1, Akira Nakagawara 1, Shigeru Sakiyama 1
PMCID: PMC5926252  PMID: 11050468

Abstract

We have recently found that DA41 exhibits marked homology with mouse PLIC‐1, PLIC‐2, frog XDRP1 and yeast DSK2. XDRP1 has been shown to be associated with cyclin A, and blocks cell division of frog embryo. In the present study, we examined the biological role(s) of DA41 in mammalian cells by overexpressing it in v‐Ha‐ras‐transformed 3Y1 cells (ras‐3Y1). Transfectants which expressed a high level of DA41 mRNA exhibited a decrease in growth rate, a reduction in saturation density, and a suppression of colony formation in soft agar medium. To clarify the molecular mechanism(s) by which DA41 affects cell growth, the effect of DA41 expression on the levels of various cell cycle‐regulatory proteins was examined. The forced expression of DA41 gene resulted in a remarkable reduction in CDK2 activity, while the amount of CDK2 did not change. These observations indicate that DA41 is involved in cell cycle regulation in ras‐3Y1 cells.

Keywords: DA41, Growth suppression, ras, 3Y1

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REFERENCES

  • 1. ) Ozaki , T. and Sakiyama , S.Tumor‐suppressive activity of N03 gene product in v‐src‐transformed rat 3Y1 fibroblasts . Cancer Res. , 54 , 646 – 648 ( 1994. ). [PubMed] [Google Scholar]
  • 2. ) Ozaki , T. , Nakamura , Y. , Enomoto , H. , Hirose , M. and Sakiyama , S.Overexpression of DAN gene product in normal rat fibroblasts causes retardation of the entry into the S phase . Cancer Res. , 55 , 895 – 900 ( 1995. ). [PubMed] [Google Scholar]
  • 3. ) Nakamura , Y. , Ozaki , T. , Nakagawara , A. and Sakiyama , S.A product of DAN, a novel candidate tumor suppressor gene, is secreted into culture medium and suppresses DNA synthesis . Eur. J. Cancer , 33 , 1986 – 1990 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 4. ) Stanley , E. , Biben , C. , Kotecha , S. , Fabri , L. , Tajbakhsh , S. , Wang , C.‐C. , Hatzistavrou , T. , Roberts , B. , Drinkwater , C. , Lah , M. , Buckingham , M. , Hilton , D. , Nash , A. , Mohun , T. and Harvey , R. P.DAN is a secreted glycoprotein related to Xenopus cerberus . Mech. Dev. , 77 , 173 – 184 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Hsu , D. R. , Economides , A. N. , Wang , X. , Eimon , P. M. and Harland , R. M.The Xenopus dorsalizing factor Gremlin identifies a novel family of secreted proteins that antagonize BMP activities . Mol. Cell , 1 , 673 – 683 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Ozaki , T. , Hishiki , T. , Toyama , Y. , Yuasa , S. , Nakagawara , A. and Sakiyama , S.Identification of a new cellular protein which can interact specifically with DAN . DNA Cell Biol. , 16 , 985 – 991 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Hanaoka , E. , Ozaki , T. , Ohira , M. , Nakamura , Y. , Suzuki , M. , Takahashi , E. , Moriya , H. , Nakagawara , A. and Sakiyama , S.Molecular cloning and expression analysis of the human DA41 gene and its mapping to chromosome 9q21.2‐q21.3 . J. Hum. Genet. , 45 , 188 – 191 ( 2000. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Wu , A.‐L. , Wang , J. , Zheleznyak , A. and Brown , E. J.Ubiquitin‐related proteins regulate interaction of vimentin intermediate filaments with the plasma membrane . Mol. Cell , 4 , 619 – 625 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Funakoshi , M. , Geley , S. , Hunt , T. , Nishimoto , T. and Kobayashi , H.Identification of XDRP1; a Xenopus protein related to yeast Dsk2p binds to the N‐terminus of cyclin A and inhibits its degradation . EMBO J. , 18 , 5009 – 5018 ( 1999. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10. ) Biggins , S. , Ivanovska , I. and Rose , M. D.Yeast ubiquitinlike genes are involved in duplication of the microtubule organizing center . J. Cell Biol. , 133 , 1331 – 1346 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. ) Chen , C. and Okayama , H.High‐efficiency transformation of mammalian cells by plasmid DNA . Mol. Cell. Biol. , 7 , 2745 – 2752 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. ) Sambrook , J. , Fritsch , E. F. and Maniatis , T. “ Molecular Cloning: A Laboratory Manual ,” 2nd Ed. ( 1989. ). Cold Spring Harbor Laboratory Press; , New York . [Google Scholar]
  • 13. ) Tate , E. H. , Wilder , M. E. , Cram , S. and Wharton , W. A.A method for staining 3T3 cell nuclei with propidium iodide in hypotonic solution . Cytometry , 4 , 211 – 215 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 14. ) Laemmli , U. K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature , 227 , 680 – 685 ( 1970. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Akiyama , T. , Ohuchi , T. , Sumida , S. , Matsumoto , K. and Toyoshima , K.Phosphorylation of the retinoblastoma protein by cdk2 . Proc. Natl. Acad. Sci. USA , 89 , 7900 – 7904 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. ) Norbury , C. and Nurse , P.Animal cell cycles and their control . Annu. Rev. Biochem. , 61 , 441 – 470 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 17. ) Draetta , G. , Luca , F. , Westendorf , J. , Brizuela , L. , Ruderman , J. and Beach , D.cdc2 protein kinase is complexed with both cyclin A and B: evidence for proteolytic inactivation of MPF . Cell , 56 , 829 – 838 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Pines , J. and Hunter , T.Isolation of a human cyclin cDNA: evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction with p34cdc2 . Cell , 58 , 833 – 846 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 19. ) Solomon , M. J. , Lee , T. and Kirschner , M. W.Role of phosphorylation in p34cdc2 activation: identification of an activating kinase . Mol. Biol. Cell , 3 , 13 – 27 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. ) Sherr , C. J. and Roberts , J. M.Inhibitors of mammalian G1 cyclin‐dependent kinases . Genes Dev. , 9 , 1149 – 1163 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Sherr , C. J.Mammalian G1 cyclins . Cell , 73 , 1059 – 1065 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Blain , S. W. , Montalvo , E. and Massague , J.Differential interaction of the cyclin‐dependent kinase (Cdk) inhibitor p27Kip1 with cyclin A‐Cdk2 and cyclin D2‐Cdk4 . J. Biol. Chem. , 272 , 25863 – 25872 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Makela , T. P. , Tassan , J.‐P. , Nigg , E. A. , Frutiger , S. , Hughes , G. J. and Weinberg , R. A.A cyclin associated with the CDβ‐activating kinase MO15 . Nature , 371 , 254 – 257 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 24. ) Chen , G. and Hitomi , M.Dissociation of CDK2 from cyclin A in response to the topoisomerase II inhibitor etoposide in v‐src‐transformed but not normal NIH3T3 cells . Exp. Cell Res. , 249 , 327 – 336 ( 1999. ). [DOI] [PubMed] [Google Scholar]
  • 25. ) Sudakin , V. , Ganoth , D. , Dahan , A. , Heller , H. , Hershko , J. , Luca , F. C. , Ruderman , J. V. and Hershko , A.The cyclosome, a large complex containing cyclin‐selective ubiquitin ligase activity, targets cyclins for destruction at the end of mitosis . Mol. Biol. Cell , 6 , 185 – 198 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

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