Mouse cells contain two distinct ras gene mRNA species that can be translated into a p21 onc protein. (original) (raw)

Mol Cell Biol. 1982 Nov; 2(11): 1339–1345.

Abstract

The Kirsten (Ki) and Harvey (Ha) strains of murine sarcoma virus encode a 21,000-dalton protein (p21 ras) which is the product of the transforming gene of these viruses. Normal cells express low levels of p21 ras encoded by cellular genes (Ki-ras and Ha-ras) homologous to the Ki and Ha murine sarcoma virus transformation genes. A bone marrow-derived mouse cell line, 416B, has been shown to express unusually high levels of p21 ras. In this manuscript, we investigated the molecular biology of p21 ras gene expression in 416B and other normal mouse cells. We identified four distinct polyadenylated and polysome-associated RNAs, two related to Ki-ras and two to Ha-ras. The levels in 416B cells of the two Ki-ras RNAs, sized 5.2 and 2.0 kilobases, were both elevated approximately 25-fold over levels found in normal mouse cells; there was no corresponding change in 416B cells in the levels of the two Ha-ras RNAs. We partially purified the two Ki-ras mRNAs and separated them by velocity sedimentation in sucrose density gradients. Both the 5.2- and 2.0-kilobase mRNAs could be translated in vitro into p21 ras. These results show that a cellular onc protein can be translated from two distinct cellular mRNA species.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.4M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

Andersen PR, Devare SG, Tronick SR, Ellis RW, Aaronson SA, Scolnick EM. Generation of BALB-MuSV and Ha-MuSC by type C virus transduction of homologous transforming genes from different species. Cell. 1981 Oct;26(1 Pt 1):129–134. [PubMed] [Google Scholar]
Bishop JM. Enemies within: the genesis of retrovirus oncogenes. Cell. 1981 Jan;23(1):5–6. [PubMed] [Google Scholar]
Chang EH, Gonda MA, Ellis RW, Scolnick EM, Lowy DR. Human genome contains four genes homologous to transforming genes of Harvey and Kirsten murine sarcoma viruses. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4848–4852. [PMC free article] [PubMed] [Google Scholar]
DeFeo D, Gonda MA, Young HA, Chang EH, Lowy DR, Scolnick EM, Ellis RW. Analysis of two divergent rat genomic clones homologous to the transforming gene of Harvey murine sarcoma virus. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3328–3332. [PMC free article] [PubMed] [Google Scholar]
Der CJ, Krontiris TG, Cooper GM. Transforming genes of human bladder and lung carcinoma cell lines are homologous to the ras genes of Harvey and Kirsten sarcoma viruses. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3637–3640. [PMC free article] [PubMed] [Google Scholar]
Dexter TM, Allen TD, Scott D, Teich NM. Isolation and characterisation of a bipotential haematopoietic cell line. Nature. 1979 Feb 8;277(5696):471–474. [PubMed] [Google Scholar]
Dhar R, Ellis RW, Shih TY, Oroszlan S, Shapiro B, Maizel J, Lowy D, Scolnick E. Nucleotide sequence of the p21 transforming protein of Harvey murine sarcoma virus. Science. 1982 Sep 3;217(4563):934–936. [PubMed] [Google Scholar]
Ellis RW, DeFeo D, Maryak JM, Young HA, Shih TY, Chang EH, Lowy DR, Scolnick EM. Dual evolutionary origin for the rat genetic sequences of Harvey murine sarcoma virus. J Virol. 1980 Nov;36(2):408–420. [PMC free article] [PubMed] [Google Scholar]
Ellis RW, Defeo D, Shih TY, Gonda MA, Young HA, Tsuchida N, Lowy DR, Scolnick EM. The p21 src genes of Harvey and Kirsten sarcoma viruses originate from divergent members of a family of normal vertebrate genes. Nature. 1981 Aug 6;292(5823):506–511. [PubMed] [Google Scholar]
Eva A, Robbins KC, Andersen PR, Srinivasan A, Tronick SR, Reddy EP, Ellmore NW, Galen AT, Lautenberger JA, Papas TS, et al. Cellular genes analogous to retroviral onc genes are transcribed in human tumour cells. Nature. 1982 Jan 14;295(5845):116–119. [PubMed] [Google Scholar]
Furth ME, Davis LJ, Fleurdelys B, Scolnick EM. Monoclonal antibodies to the p21 products of the transforming gene of Harvey murine sarcoma virus and of the cellular ras gene family. J Virol. 1982 Jul;43(1):294–304. [PMC free article] [PubMed] [Google Scholar]
Hayward WS, Neel BG, Astrin SM. Activation of a cellular onc gene by promoter insertion in ALV-induced lymphoid leukosis. Nature. 1981 Apr 9;290(5806):475–480. [PubMed] [Google Scholar]
Heilig R, Perrin F, Gannon F, Mandel JL, Chambon P. The ovalbumin gene family: structure of the X gene and evolution of duplicated split genes. Cell. 1980 Jul;20(3):625–637. [PubMed] [Google Scholar]
Lehrach H, Diamond D, Wozney JM, Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. [PubMed] [Google Scholar]
Parada LF, Tabin CJ, Shih C, Weinberg RA. Human EJ bladder carcinoma oncogene is homologue of Harvey sarcoma virus ras gene. Nature. 1982 Jun 10;297(5866):474–478. [PubMed] [Google Scholar]
Payne GS, Bishop JM, Varmus HE. Multiple arrangements of viral DNA and an activated host oncogene in bursal lymphomas. Nature. 1982 Jan 21;295(5846):209–214. [PubMed] [Google Scholar]
Ross J. A precursor of globin messenger RNA. J Mol Biol. 1976 Sep 15;106(2):403–420. [PubMed] [Google Scholar]
Ruscetti S, Troxler D, Linemeyer D, Scolnick E. Three laboratory strains of spleen focus-forming virus: comparison of their genomes and translational products. J Virol. 1980 Jan;33(1):140–151. [PMC free article] [PubMed] [Google Scholar]
Scolnick EM, Weeks MO, Shih TY, Ruscetti SK, Dexter TM. Markedly elevated levels of an endogenous sarc protein in a hemopoietic precursor cell line. Mol Cell Biol. 1981 Jan;1(1):66–74. [PMC free article] [PubMed] [Google Scholar]
Scolnick EM, Williams D, Maryak J, Vass W, Goldberg RJ, Parks WP. Type C particle-positive and type C particle-negative rat cell lines: characterization of the coding capacity of endogenous sarcoma virus-specific RNA. J Virol. 1976 Dec;20(3):570–582. [PMC free article] [PubMed] [Google Scholar]
Setzer DR, McGrogan M, Nunberg JH, Schimke RT. Size heterogeneity in the 3' end of dihydrofolate reductase messenger RNAs in mouse cells. Cell. 1980 Nov;22(2 Pt 2):361–370. [PubMed] [Google Scholar]
Shih TY, Weeks MO, Gruss P, Dhar R, Oroszlan S, Scolnick EM. Identification of a precursor in the biosynthesis of the p21 transforming protein of harvey murine sarcoma virus. J Virol. 1982 Apr;42(1):253–261. [PMC free article] [PubMed] [Google Scholar]
Shih TY, Weeks MO, Young HA, Scholnick EM. Identification of a sarcoma virus-coded phosphoprotein in nonproducer cells transformed by Kirsten or Harvey murine sarcoma virus. Virology. 1979 Jul 15;96(1):64–79. [PubMed] [Google Scholar]
Tsuchida N, Ryder T, Ohtsubo E. Nucleotide sequence of the oncogene encoding the p21 transforming protein of Kirsten murine sarcoma virus. Science. 1982 Sep 3;217(4563):937–939. [PubMed] [Google Scholar]
Vennström B, Bishop JM. Isolation and characterization of chicken DNA homologous to the two putative oncogenes of avian erythroblastosis virus. Cell. 1982 Jan;28(1):135–143. [PubMed] [Google Scholar]
Wahl GM, Stern M, Stark GR. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis