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. 1988 Jan;62(1):349–353. doi: 10.1128/jvi.62.1.349-353.1988

Avian sarcoma-leukosis virus pol-endo proteins expressed independently in mammalian cells accumulate in the nucleus but can be directed to other cellular compartments.

C Morris-Vasios 1, J P Kochan 1, A M Skalka 1
PMCID: PMC250537  PMID: 2446017

Abstract

Eucaryotic expression vectors have been used to study transient expression of the avian sarcoma-leukosis retrovirus pol-endo protein in COS cells. The constructs encode proteins with N termini identical to that of authentic viral pp32 endonuclease with the exception of a single met residue encoded by the initiator AUG. The C termini correspond to unprocessed viral pol protein, authentic processed pp32, or a derivative which includes eight amino acids from the unprocessed portion. All three proteins localize to the nucleus. However, when the pol-endo domain is fused to a secretory signal peptide, the protein is found in medium and appears also to localize in the Golgi bodies and the cell membrane. These and derivative vectors will make it possible to assess the consequence of retroviral pol gene expression in eucaryotic cells.

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Selected References

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  1. Alexander F., Leis J., Soltis D. A., Crowl R. M., Danho W., Poonian M. S., Pan Y. C., Skalka A. M. Proteolytic processing of avian sarcoma and leukosis viruses pol-endo recombinant proteins reveals another pol gene domain. J Virol. 1987 Feb;61(2):534–542. doi: 10.1128/jvi.61.2.534-542.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Challberg M. D., Kelly T. J., Jr Adenovirus DNA replication in vitro. Proc Natl Acad Sci U S A. 1979 Feb;76(2):655–659. doi: 10.1073/pnas.76.2.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chiu I. M., Callahan R., Tronick S. R., Schlom J., Aaronson S. A. Major pol gene progenitors in the evolution of oncoviruses. Science. 1984 Jan 27;223(4634):364–370. doi: 10.1126/science.6197754. [DOI] [PubMed] [Google Scholar]
  4. Cullen B. R., Lomedico P. T., Ju G. Transcriptional interference in avian retroviruses--implications for the promoter insertion model of leukaemogenesis. Nature. 1984 Jan 19;307(5948):241–245. doi: 10.1038/307241a0. [DOI] [PubMed] [Google Scholar]
  5. Cullen B. R. Use of eukaryotic expression technology in the functional analysis of cloned genes. Methods Enzymol. 1987;152:684–704. doi: 10.1016/0076-6879(87)52074-2. [DOI] [PubMed] [Google Scholar]
  6. Curran T., Miller A. D., Zokas L., Verma I. M. Viral and cellular fos proteins: a comparative analysis. Cell. 1984 Feb;36(2):259–268. doi: 10.1016/0092-8674(84)90219-8. [DOI] [PubMed] [Google Scholar]
  7. DeChiara T. M., Erlitz F., Tarnowski S. J. Procedures for in vitro DNA mutagenesis of human leukocyte interferon sequences. Methods Enzymol. 1986;119:403–415. doi: 10.1016/0076-6879(86)19060-4. [DOI] [PubMed] [Google Scholar]
  8. Donehower L. A., Varmus H. E. A mutant murine leukemia virus with a single missense codon in pol is defective in a function affecting integration. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6461–6465. doi: 10.1073/pnas.81.20.6461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hippenmeyer P. J., Grandgenett D. P. Requirement of the avian retrovirus pp32 DNA binding protein domain for replication. Virology. 1984 Sep;137(2):358–370. doi: 10.1016/0042-6822(84)90228-9. [DOI] [PubMed] [Google Scholar]
  10. Johnson M. S., McClure M. A., Feng D. F., Gray J., Doolittle R. F. Computer analysis of retroviral pol genes: assignment of enzymatic functions to specific sequences and homologies with nonviral enzymes. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7648–7652. doi: 10.1073/pnas.83.20.7648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Lingappa V. R., Chaidez J., Yost C. S., Hedgpeth J. Determinants for protein localization: beta-lactamase signal sequence directs globin across microsomal membranes. Proc Natl Acad Sci U S A. 1984 Jan;81(2):456–460. doi: 10.1073/pnas.81.2.456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Panganiban A. T., Temin H. M. The retrovirus pol gene encodes a product required for DNA integration: identification of a retrovirus int locus. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7885–7889. doi: 10.1073/pnas.81.24.7885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Schwartzberg P., Colicelli J., Gordon M. L., Goff S. P. Mutations in the gag gene of Moloney murine leukemia virus: effects on production of virions and reverse transcriptase. J Virol. 1984 Mar;49(3):918–924. doi: 10.1128/jvi.49.3.918-924.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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