Abstract
A vaccinia virus host range mutant (hr mutant) deleted of 18 kilobase pairs at the left end of the genome has been employed to precisely map a viral gene required for multiplication in human cells. DNA fragments from the wild-type virus were inserted into the thymidine kinase gene of mutant virus by means of in vivo homologous recombination, and the recombinants obtained were screened for their ability to multiply in human cells. A short sequence, 855 base pairs long, overlapping the HindIII M and K fragments was able to restore a normal host range on the mutant virus. A single long open reading frame that could encode a polypeptide of 32.5 kDa was found in the nucleotide sequence of the host range gene. The direction of transcription and the length of the open reading frame are in excellent agreement with previous mapping of mRNAs within this region of the genome. In vitro translation of infected cell early mRNA, selected by hybridization to the host range gene, yielded a prominent polypeptide product whose size (29 kDa) was close to that expected from the predicted amino acid sequence. The phenotype of the hr mutant suggests that the host range gene plays a role in maintaining a high level of protein synthesis in human cells. It may behave positively by complementing the lack of an analogous cellular activity or negatively by antagonizing a cell function that inhibits viral multiplication.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Archard L. C., Mackett M., Barnes D. E., Dumbell K. R. The genome structure of cowpox virus white pock variants. J Gen Virol. 1984 May;65(Pt 5):875–886. doi: 10.1099/0022-1317-65-5-875. [DOI] [PubMed] [Google Scholar]
- Archard L. C., Mackett M. Restriction endonuclease analysis of red cowpox virus and its white pock variant. J Gen Virol. 1979 Oct;45(1):51–63. doi: 10.1099/0022-1317-45-1-51. [DOI] [PubMed] [Google Scholar]
- Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blomquist M. C., Hunt L. T., Barker W. C. Vaccinia virus 19-kilodalton protein: relationship to several mammalian proteins, including two growth factors. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7363–7367. doi: 10.1073/pnas.81.23.7363. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Boone R. F., Moss B. Sequence complexity and relative abundance of vaccinia virus mRNA's synthesized in vivo and in vitro. J Virol. 1978 Jun;26(3):554–569. doi: 10.1128/jvi.26.3.554-569.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown J. P., Twardzik D. R., Marquardt H., Todaro G. J. Vaccinia virus encodes a polypeptide homologous to epidermal growth factor and transforming growth factor. Nature. 1985 Feb 7;313(6002):491–492. doi: 10.1038/313491a0. [DOI] [PubMed] [Google Scholar]
- Chen H. R., Barker W. C. Similarity of vaccinia 28K, v-erb-B and EGF receptors. Nature. 1985 Jul 18;316(6025):219–220. doi: 10.1038/316219b0. [DOI] [PubMed] [Google Scholar]
- Condit R. C., Motyczka A., Spizz G. Isolation, characterization, and physical mapping of temperature-sensitive mutants of vaccinia virus. Virology. 1983 Jul 30;128(2):429–443. doi: 10.1016/0042-6822(83)90268-4. [DOI] [PubMed] [Google Scholar]
- Cooper J. A., Wittek R., Moss B. Hybridization selection and cell-free translation of mRNA's encoded within the inverted terminal repetition of the vaccinia virus genome. J Virol. 1981 Jan;37(1):284–294. doi: 10.1128/jvi.37.1.284-294.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drillien R., Koehren F., Kirn A. Host range deletion mutant of vaccinia virus defective in human cells. Virology. 1981 Jun;111(2):488–499. doi: 10.1016/0042-6822(81)90351-2. [DOI] [PubMed] [Google Scholar]
- Drillien R., Spehner D. Physical mapping of vaccinia virus temperature-sensitive mutations. Virology. 1983 Dec;131(2):385–393. doi: 10.1016/0042-6822(83)90506-8. [DOI] [PubMed] [Google Scholar]
- Dumbell K. R., Archard L. C. Comparison of white pock (h) mutants of monkeypox virus with parental monkeypox and with variola-like viruses isolated from animals. Nature. 1980 Jul 3;286(5768):29–32. doi: 10.1038/286029a0. [DOI] [PubMed] [Google Scholar]
- Ensinger M. J., Rovinsky M. Marker rescue of temperature-sensitive mutations of vaccinia virus WR: correlation of genetic and physical maps. J Virol. 1983 Nov;48(2):419–428. doi: 10.1128/jvi.48.2.419-428.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Esposito J. J., Cabradilla C. D., Nakano J. H., Obijeski J. F. Intragenomic sequence transposition in monkeypox virus. Virology. 1981 Mar;109(2):231–243. doi: 10.1016/0042-6822(81)90495-5. [DOI] [PubMed] [Google Scholar]
- Gillard S., Spehner D., Drillien R. Mapping of a vaccinia host range sequence by insertion into the viral thymidine kinase gene. J Virol. 1985 Jan;53(1):316–318. doi: 10.1128/jvi.53.1.316-318.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
- Isle H. B., Venkatesan S., Moss B. Cell-free translation of early and late mRNAs selected by hybridization to cloned DNA fragments derived from the left 14 million to 72 million daltons of the vaccinia virus genome. Virology. 1981 Jul 15;112(1):306–317. doi: 10.1016/0042-6822(81)90636-x. [DOI] [PubMed] [Google Scholar]
- Kieny M. P., Lathe R., Drillien R., Spehner D., Skory S., Schmitt D., Wiktor T., Koprowski H., Lecocq J. P. Expression of rabies virus glycoprotein from a recombinant vaccinia virus. Nature. 1984 Nov 8;312(5990):163–166. doi: 10.1038/312163a0. [DOI] [PubMed] [Google Scholar]
- Kieny M. P., Lathe R., Lecocq J. P. New versatile cloning and sequencing vectors based on bacteriophage M13. Gene. 1983 Dec;26(1):91–99. doi: 10.1016/0378-1119(83)90039-2. [DOI] [PubMed] [Google Scholar]
- Kwoh T. J., Engler J. A. The nucleotide sequence of the chicken thymidine kinase gene and the relationship of its predicted polypeptide to that of the vaccinia virus thymidine kinase. Nucleic Acids Res. 1984 May 11;12(9):3959–3971. doi: 10.1093/nar/12.9.3959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lake J. R., Cooper P. D. Deletions of the terminal sequences in the genomes of the white pock (u) and host-restricted (p) mutants of rabbitpox virus. J Gen Virol. 1980 May;48(1):135–147. doi: 10.1099/0022-1317-48-1-135. [DOI] [PubMed] [Google Scholar]
- Mackett M., Archard L. C. Conservation and variation in Orthopoxvirus genome structure. J Gen Virol. 1979 Dec;45(3):683–701. doi: 10.1099/0022-1317-45-3-683. [DOI] [PubMed] [Google Scholar]
- Mackett M., Smith G. L., Moss B. Vaccinia virus: a selectable eukaryotic cloning and expression vector. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7415–7419. doi: 10.1073/pnas.79.23.7415. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McFadden G., Dales S. Biogenesis of poxviruses: mirror-image deletions in vaccinia virus DNA. Cell. 1979 Sep;18(1):101–108. doi: 10.1016/0092-8674(79)90358-1. [DOI] [PubMed] [Google Scholar]
- Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
- Morgan J. R., Roberts B. E. Organization of RNA transcripts from a vaccinia virus early gene cluster. J Virol. 1984 Aug;51(2):283–297. doi: 10.1128/jvi.51.2.283-297.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moss B., Winters E., Cooper J. A. Deletion of a 9,000-base-pair segment of the vaccinia virus genome that encodes nonessential polypeptides. J Virol. 1981 Nov;40(2):387–395. doi: 10.1128/jvi.40.2.387-395.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moyer R. W., Graves R. L., Rothe C. T. The white pock (mu) mutants of rabbit poxvirus. III. Terminal DNA sequence duplication and transposition in rabbit poxvirus. Cell. 1980 Nov;22(2 Pt 2):545–553. doi: 10.1016/0092-8674(80)90364-5. [DOI] [PubMed] [Google Scholar]
- Moyer R. W., Rothe C. T. The white pock mutants of rabbit poxvirus. I. Spontaneous host range mutants contain deletions. Virology. 1980 Apr 15;102(1):119–132. doi: 10.1016/0042-6822(80)90075-6. [DOI] [PubMed] [Google Scholar]
- Panicali D., Davis S. W., Mercer S. R., Paoletti E. Two major DNA variants present in serially propagated stocks of the WR strain of vaccinia virus. J Virol. 1981 Mar;37(3):1000–1010. doi: 10.1128/jvi.37.3.1000-1010.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
- Pickup D. J., Ink B. S., Parsons B. L., Hu W., Joklik W. K. Spontaneous deletions and duplications of sequences in the genome of cowpox virus. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6817–6821. doi: 10.1073/pnas.81.21.6817. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reisner A. H. Similarity between the vaccinia virus 19K early protein and epidermal growth factor. 1985 Feb 28-Mar 6Nature. 313(6005):801–803. doi: 10.1038/313801a0. [DOI] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [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]
- Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
- Weir J. P., Moss B. Regulation of expression and nucleotide sequence of a late vaccinia virus gene. J Virol. 1984 Sep;51(3):662–669. doi: 10.1128/jvi.51.3.662-669.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]