Abstract
A series of deletions and insertions utilizing the herpesvirus thymidine kinase gene (tk) were constructed in the murine retrovirus Friend spleen focus-forming virus (SFFV). In all cases, the coding region for the SFFV-specific glycoprotein (gp55), which is implicated in erythroleukemic transformation, was left intact. These SFFV-TK and SFFV deletion vectors were analyzed for expression of tk and gp55 after DNA-mediated gene transfer. In addition, virus rescued by cotransfection of these vectors with Moloney murine leukemia virus was analyzed for infectious TK-transducing virus, gp55 expression, and erythroleukemia-inducing ability. The experiments demonstrated that deletions or insertions within the intron for the gp55 env gene can interfere with expression of gp55 after both DNA-mediated gene transfer and virus infection. In contrast, the gene transfer efficiency of the tk gene was unaffected in the SFFV-TK vectors, and high-titer infectious TK virus could be recovered. Revertant viruses capable of inducing erythroleukemia and expressing gp55 were generated after cotransfection of the SFFV-TK vectors with murine leukemia virus. The revertant viruses lost both tk sequences and the ability to transduce TK- fibroblasts to a TK+ phenotype. These experiments demonstrate that segregation of the TK and erythroleukemia functions can occur in retrovirus vectors which initially carry both markers.
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Selected References
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- AXELRAD A. A., STEEVES R. A. ASSAY FOR FRIEND LEUKEMIA VIRUS: RAPID QUANTITATIVE METHOD BASED ON ENUMERATION OF MACROSCOPIC SPLEEN FOCI IN MICE. Virology. 1964 Nov;24:513–518. doi: 10.1016/0042-6822(64)90199-0. [DOI] [PubMed] [Google Scholar]
- Bernstein A., Mak T. W., Stephenson J. R. The Friend virus genome: evidence for the stable association of MuLV sequences and sequences involved in erythroleukemic transformation. Cell. 1977 Sep;12(1):287–294. doi: 10.1016/0092-8674(77)90206-9. [DOI] [PubMed] [Google Scholar]
- Bilello J. A., Colletta G., Warnecke G., Koch G., Frisby D., Pragnell I. B., Ostertag W. Analysis of the expression of spleen focus-forming virus (SFFV)-related RNA and gp55, a Friend and Rauscher virus-specific protein. Virology. 1980 Dec;107(2):331–344. doi: 10.1016/0042-6822(80)90301-3. [DOI] [PubMed] [Google Scholar]
- Bosselman R. A., Van Griensven L. J., Vogt M., Verma I. M. Genome organization of retroviruses IX. Analysis of the genomes of Friend spleen focus-forming (F-SFFV) and helper murine leukemia viruses by heteroduplex-formation. Virology. 1980 Apr 15;102(1):234–239. doi: 10.1016/0042-6822(80)90088-4. [DOI] [PubMed] [Google Scholar]
- Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
- Brugge J. S., Erikson R. L. Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature. 1977 Sep 22;269(5626):346–348. doi: 10.1038/269346a0. [DOI] [PubMed] [Google Scholar]
- Chen I. S., Temin H. M. Substitution of 5' helper virus sequences into non-rel portion of reticuloendotheliosis virus strain T suppresses transformation of chicken spleen cells. Cell. 1982 Nov;31(1):111–120. doi: 10.1016/0092-8674(82)90410-x. [DOI] [PubMed] [Google Scholar]
- Clark S. P., Mak T. W. Complete nucleotide sequence of an infectious clone of Friend spleen focus-forming provirus: gp55 is an envelope fusion glycoprotein. Proc Natl Acad Sci U S A. 1983 Aug;80(16):5037–5041. doi: 10.1073/pnas.80.16.5037. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
- Devare S. G., Reddy E. P., Robbins K. C., Andersen P. R., Tronick S. R., Aaronson S. A. Nucleotide sequence of the transforming gene of simian sarcoma virus. Proc Natl Acad Sci U S A. 1982 May;79(10):3179–3182. doi: 10.1073/pnas.79.10.3179. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Edgell M. H., Polsky F. I. Use of preparative gel electrophoresis for DNA fragment isolation. Methods Enzymol. 1980;65(1):319–327. doi: 10.1016/s0076-6879(80)65042-3. [DOI] [PubMed] [Google Scholar]
- Enquist L. W., Vande Woude G. F., Wagner M., Smiley J. R., Summers W. C. Construction and characterization of a recombinant plasmid encoding the gene for the thymidine kinase of Herpes simplex type 1 virus. Gene. 1979 Nov;7(3-4):335–342. doi: 10.1016/0378-1119(79)90052-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Harbers K., Schnieke A., Stuhlmann H., Jähner D., Jaenisch R. DNA methylation and gene expression: endogenous retroviral genome becomes infectious after molecular cloning. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7609–7613. doi: 10.1073/pnas.78.12.7609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Joyner A. L., Bernstein A. Retrovirus transduction: generation of infectious retroviruses expressing dominant and selectable genes is associated with in vivo recombination and deletion events. Mol Cell Biol. 1983 Dec;3(12):2180–2190. doi: 10.1128/mcb.3.12.2180. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Joyner A., Yamamoto Y., Bernstein A. Retrovirus long terminal repeats activate expression of coding sequences for the herpes simplex virus thymidine kinase gene. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1573–1577. doi: 10.1073/pnas.79.5.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klempnauer K. H., Gonda T. J., Bishop J. M. Nucleotide sequence of the retroviral leukemia gene v-myb and its cellular progenitor c-myb: the architecture of a transduced oncogene. Cell. 1982 Dec;31(2 Pt 1):453–463. doi: 10.1016/0092-8674(82)90138-6. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Linemeyer D. L., Menke J. G., Ruscetti S. K., Evans L. H., Scolnick E. M. Envelope gene sequences which encode the gp52 protein of spleen focus-forming virus are required for the induction of erythroid cell proliferation. J Virol. 1982 Jul;43(1):223–233. doi: 10.1128/jvi.43.1.223-233.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Linemeyer D. L., Ruscetti S. K., Scolnick E. M., Evans L. H., Duesberg P. H. Biological activity of the spleen focus-forming virus is encoded by a molecularly cloned subgenomic fragment of spleen focus-forming virus DNA. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1401–1405. doi: 10.1073/pnas.78.3.1401. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacDonald M. E., Reynolds F. H., Jr, Van de Ven W. J., Stephenson J. R., Mak T. W., Bernstein A. Anemia- and polycythemia-inducing isolates of Friend spleen focus-forming virus. Biological and molecular evidence for two distinct viral genomes. J Exp Med. 1980 Jun 1;151(6):1477–1492. doi: 10.1084/jem.151.6.1477. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mager D. L., Mak T. W., Bernstein A. Quantitative colony method for tumorigenic cells transformed by two distinct strains of Friend leukemia virus. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1703–1707. doi: 10.1073/pnas.78.3.1703. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mager D., MacDonald M. E., Robson I. B., Mak T. W., Bernstein A. Clonal analysis of the late stages of erythroleukemia induced by two distinct strains of Friend leukemia virus. Mol Cell Biol. 1981 Aug;1(8):721–730. doi: 10.1128/mcb.1.8.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Papas T. S., Rushlow K. E., Watson D. K., Lautenberger J. A., Perbal B., Baluda M. E., Reddy E. P. Nucleotide sequence and organization of the transforming region and large terminal redundancies (LTR) of avian myeloblastosis virus (AMV). J Cell Biochem. 1982;20(2):95–103. doi: 10.1002/jcb.240200202. [DOI] [PubMed] [Google Scholar]
- Racevskis J., Koch G. Viral protein synthesis in Friend erythroleukemia cell lines. J Virol. 1977 Jan;21(1):328–337. doi: 10.1128/jvi.21.1.328-337.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reddy E. P., Smith M. J., Aaronson S. A. Complete nucleotide sequence and organization of the Moloney murine sarcoma virus genome. Science. 1981 Oct 23;214(4519):445–450. doi: 10.1126/science.6170110. [DOI] [PubMed] [Google Scholar]
- Rowe W. P., Pugh W. E., Hartley J. W. Plaque assay techniques for murine leukemia viruses. Virology. 1970 Dec;42(4):1136–1139. doi: 10.1016/0042-6822(70)90362-4. [DOI] [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. doi: 10.1128/jvi.33.1.140-151.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
- Varmus H. E., Quintrell N., Ortiz S. Retroviruses as mutagens: insertion and excision of a nontransforming provirus alter expression of a resident transforming provirus. Cell. 1981 Jul;25(1):23–36. doi: 10.1016/0092-8674(81)90228-2. [DOI] [PubMed] [Google Scholar]
- Wahl G. M., Stern M., Stark G. R. 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. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wendling F., Moreau-Gachelin F., Tambourin P. Emergence of tumorigenic cells during the course of Friend virus leukemias. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3614–3618. doi: 10.1073/pnas.78.6.3614. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]
- Yamamoto Y., Gamble C. L., Clark S. P., Joyner A., Shibuya T., MacDonald M. E., Mager D., Bernstein A., Mak T. W. Clonal analysis of early and late stages of erythroleukemia induced by molecular clones of integrated spleen focus-forming virus. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6893–6897. doi: 10.1073/pnas.78.11.6893. [DOI] [PMC free article] [PubMed] [Google Scholar]