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. 1992 Apr 11;20(7):1637–1642. doi: 10.1093/nar/20.7.1637

A single amino acid change restores DNA cytosine methyltransferase activity in a cloned chlorella virus pseudogene.

Y Zhang 1, M Nelson 1, J L Van Etten 1
PMCID: PMC312249  PMID: 1579454

Abstract

The chlorella virus PBCV-1 contains an open reading frame, named P17-ORF4, which differs by eight amino acids from a DNA cytosine methyltransferase, M.CviJI, encoded by a different chlorella virus IL-3A. Whereas IL-3A expresses M.CviJI, which methylates the central cytosine in (A/G)GC(T/C/G) sequences, P17-ORF4 is non-functional. Gene fusions between P17-ORF4 and M.CviJI and site-directed point mutations revealed that changing Gln188 to Lys188 abolishes M.CviJI methyltransferase activity. Conversely, changing Lys188 in P17-ORF4 to Gln188 results in M.CviJI activity. The other altered seven amino acids do not appear to affect M.CviJI activity.

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Selected References

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  1. Burbank D. E., Shields S. L., Schuster A. M., Van Etten J. L. 5-Azacytidine-resistant mutants of Chlorella virus IL-3A. Virology. 1990 May;176(1):311–315. doi: 10.1016/0042-6822(90)90261-o. [DOI] [PubMed] [Google Scholar]
  2. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  3. Kuo T. T., Huang T. C., Teng M. H. 5-Methylcytosine replacing cytosine in the deoxyribonucleic acid of a bacteriophage for Xanthomonas oryzae. J Mol Biol. 1968 Jul 14;34(2):373–375. doi: 10.1016/0022-2836(68)90263-5. [DOI] [PubMed] [Google Scholar]
  4. Lange C., Jugel A., Walter J., Noyer-Weidner M., Trautner T. A. 'Pseudo' domains in phage-encoded DNA methyltransferases. Nature. 1991 Aug 15;352(6336):645–648. doi: 10.1038/352645a0. [DOI] [PubMed] [Google Scholar]
  5. Nelson M., McClelland M. Site-specific methylation: effect on DNA modification methyltransferases and restriction endonucleases. Nucleic Acids Res. 1991 Apr 25;19 (Suppl):2045–2071. doi: 10.1093/nar/19.suppl.2045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Pósfai J., Bhagwat A. S., Pósfai G., Roberts R. J. Predictive motifs derived from cytosine methyltransferases. Nucleic Acids Res. 1989 Apr 11;17(7):2421–2435. doi: 10.1093/nar/17.7.2421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Schuster A. M., Burbank D. E., Meister B., Skrdla M. P., Meints R. H., Hattman S., Swinton D., Van Etten J. L. Characterization of viruses infecting a eukaryotic Chlorella-like green alga. Virology. 1986 Apr 15;150(1):170–177. doi: 10.1016/0042-6822(86)90276-x. [DOI] [PubMed] [Google Scholar]
  8. Schuster A. M., Girton L., Burbank D. E., Van Etten J. L. Infection of a Chlorella-like alga with the virus PBCV-1: transcriptional studies. Virology. 1986 Jan 15;148(1):181–189. doi: 10.1016/0042-6822(86)90413-7. [DOI] [PubMed] [Google Scholar]
  9. Shields S. L., Burbank D. E., Grabherr R., van Etten J. L. Cloning and sequencing the cytosine methyltransferase gene M. CviJI from Chlorella virus IL-3A. Virology. 1990 May;176(1):16–24. doi: 10.1016/0042-6822(90)90225-g. [DOI] [PubMed] [Google Scholar]
  10. Strasser P., Zhang Y. P., Rohozinski J., Van Etten J. L. The termini of the chlorella virus PBCV-1 genome are identical 2.2-kbp inverted repeats. Virology. 1991 Feb;180(2):763–769. doi: 10.1016/0042-6822(91)90089-t. [DOI] [PubMed] [Google Scholar]
  11. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Van Etten J. L., Burbank D. E., Schuster A. M., Meints R. H. Lytic viruses infecting a Chlorella-like alga. Virology. 1985 Jan 15;140(1):135–143. doi: 10.1016/0042-6822(85)90452-0. [DOI] [PubMed] [Google Scholar]
  13. Van Etten J. L., Lane L. C., Meints R. H. Viruses and viruslike particles of eukaryotic algae. Microbiol Rev. 1991 Dec;55(4):586–620. doi: 10.1128/mr.55.4.586-620.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Van Etten J. L., Schuster A. M., Girton L., Burbank D. E., Swinton D., Hattman S. DNA methylation of viruses infecting a eukaryotic Chlorella-like green alga. Nucleic Acids Res. 1985 May 24;13(10):3471–3478. doi: 10.1093/nar/13.10.3471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Warren R. A. Modified bases in bacteriophage DNAs. Annu Rev Microbiol. 1980;34:137–158. doi: 10.1146/annurev.mi.34.100180.001033. [DOI] [PubMed] [Google Scholar]
  16. Xia Y. N., Burbank D. E., Uher L., Rabussay D., Van Etten J. L. IL-3A virus infection of a Chlorella-like green alga induces a DNA restriction endonuclease with novel sequence specificity. Nucleic Acids Res. 1987 Aug 11;15(15):6075–6090. doi: 10.1093/nar/15.15.6075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Xia Y. N., Burbank D. E., Uher L., Rabussay D., Van Etten J. L. Restriction endonuclease activity induced by PBCV-1 virus infection of a Chlorella-like green alga. Mol Cell Biol. 1986 May;6(5):1430–1439. doi: 10.1128/mcb.6.5.1430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Xia Y. N., Van Etten J. L. DNA methyltransferase induced by PBCV-1 virus infection of a Chlorella-like green alga. Mol Cell Biol. 1986 May;6(5):1440–1445. doi: 10.1128/mcb.6.5.1440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zhang Y. P., Burbank D. E., Van Etten J. L. Chlorella viruses isolated in China. Appl Environ Microbiol. 1988 Sep;54(9):2170–2173. doi: 10.1128/aem.54.9.2170-2173.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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