Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1984 Oct;4(10):2103–2108. doi: 10.1128/mcb.4.10.2103

Loss of chloroplast DNA methylation during dedifferentiation of Chlamydomonas reinhardi gametes.

H Sano, C Grabowy, R Sager
PMCID: PMC369028  PMID: 6095040

Abstract

In Chlamydomonas reinhardi the chloroplast DNA (ch;DNA) of mating type plus cells undergoes cyclical methylation and demethylation during the life cycle. Methylation occurs during gametogenesis, and fully differentiated gametes can be dedifferentiated back to vegetative cells which contain nonmethylated chlDNA by the addition of a nitrogen source for growth. We examined the dedifferentiation process and found that the mating ability of gametes was lost rapidly after the start of dedifferentiation at a time when the chlDNA was still methylated. The enzymatic activity of the 200-kilodalton DNA methyltransferase was lost at a rate consistent with the rate of dilution during cell division. Methylation of chlDNA decreased at a slower rate than was expected from cell division alone but was consistent with the continuing activity of the preexisting methyltransferase so long as it was present. These results support the hypothesis that demethylation of chlDNA occurs by dilution out of enzymatic methylating activity rather than by enzymatic demethylation.

Full text

PDF
2103

Images in this article

2105Image
on p.2105

Selected References

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

  1. Adams R. L., McKay E. L., Craig L. M., Burdon R. H. Mouse DNA methylase: methylation of native DNA. Biochim Biophys Acta. 1979 Feb 27;561(2):345–357. doi: 10.1016/0005-2787(79)90143-6. [DOI] [PubMed] [Google Scholar]
  2. Bestor T. H., Ingram V. M. Two DNA methyltransferases from murine erythroleukemia cells: purification, sequence specificity, and mode of interaction with DNA. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5559–5563. doi: 10.1073/pnas.80.18.5559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolen P. L., Grant D. M., Swinton D., Boynton J. E., Gillham N. W. Extensive methylation of chloroplast DNA by a nuclear gene mutation does not affect chloroplast gene transmission in chlamydomonas. Cell. 1982 Feb;28(2):335–343. doi: 10.1016/0092-8674(82)90351-8. [DOI] [PubMed] [Google Scholar]
  4. Burton W. G., Grabowy C. T., Sager R. Role of methylation in the modification and restriction of chloroplast DNA in Chlamydomonas. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1390–1394. doi: 10.1073/pnas.76.3.1390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DOSKOCIL J., SORM F. Distribution of 5-methylcytosine in pyrimidine sequences of deoxyribonucleic acids. Biochim Biophys Acta. 1962 Jun 11;55:953–959. doi: 10.1016/0006-3002(62)90909-5. [DOI] [PubMed] [Google Scholar]
  7. Doerfler W. DNA methylation and gene activity. Annu Rev Biochem. 1983;52:93–124. doi: 10.1146/annurev.bi.52.070183.000521. [DOI] [PubMed] [Google Scholar]
  8. Ehrlich M., Wang R. Y. 5-Methylcytosine in eukaryotic DNA. Science. 1981 Jun 19;212(4501):1350–1357. doi: 10.1126/science.6262918. [DOI] [PubMed] [Google Scholar]
  9. Feinberg A. P., Vogelstein B. Hypomethylation distinguishes genes of some human cancers from their normal counterparts. Nature. 1983 Jan 6;301(5895):89–92. doi: 10.1038/301089a0. [DOI] [PubMed] [Google Scholar]
  10. Gjerset R. A., Martin D. W., Jr Presence of a DNA demethylating activity in the nucleus of murine erythroleukemic cells. J Biol Chem. 1982 Aug 10;257(15):8581–8583. [PubMed] [Google Scholar]
  11. Gruenbaum Y., Cedar H., Razin A. Substrate and sequence specificity of a eukaryotic DNA methylase. Nature. 1982 Feb 18;295(5850):620–622. doi: 10.1038/295620a0. [DOI] [PubMed] [Google Scholar]
  12. Gruenbaum Y., Naveh-Many T., Cedar H., Razin A. Sequence specificity of methylation in higher plant DNA. Nature. 1981 Aug 27;292(5826):860–862. doi: 10.1038/292860a0. [DOI] [PubMed] [Google Scholar]
  13. Harrison J. J., Anisowicz A., Gadi I. K., Raffeld M., Sager R. Azacytidine-induced tumorigenesis of CHEF/18 cells: correlated DNA methylation and chromosome changes. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6606–6610. doi: 10.1073/pnas.80.21.6606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kuo K. C., McCune R. A., Gehrke C. W., Midgett R., Ehrlich M. Quantitative reversed-phase high performance liquid chromatographic determination of major and modified deoxyribonucleosides in DNA. Nucleic Acids Res. 1980 Oct 24;8(20):4763–4776. doi: 10.1093/nar/8.20.4763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Razin A., Riggs A. D. DNA methylation and gene function. Science. 1980 Nov 7;210(4470):604–610. doi: 10.1126/science.6254144. [DOI] [PubMed] [Google Scholar]
  16. Royer H. D., Sager R. Methylation of chloroplast DNAs in the life cycle of Chlamydomonas. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5794–5798. doi: 10.1073/pnas.76.11.5794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. SAGER R., GRANICK S. Nutritional control of sexuality in Chlamydomonas reinhardi. J Gen Physiol. 1954 Jul 20;37(6):729–742. doi: 10.1085/jgp.37.6.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sager R., Grabowy C. Differential methylation of chloroplast DNA regulates maternal inheritance in a methylated mutant of Chlamydomonas. Proc Natl Acad Sci U S A. 1983 May;80(10):3025–3029. doi: 10.1073/pnas.80.10.3025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sager R., Grabowy C., Sano H. The mat-1 gene in Chlamydomonas regulates DNA methylation during gametogenesis. Cell. 1981 Apr;24(1):41–47. doi: 10.1016/0092-8674(81)90499-2. [DOI] [PubMed] [Google Scholar]
  20. Sager R., Sano H., Grabowy C. T. Control of maternal inheritance by DNA methylation in chlamydomonas. Curr Top Microbiol Immunol. 1984;108:157–173. doi: 10.1007/978-3-642-69370-0_11. [DOI] [PubMed] [Google Scholar]
  21. Sano H., Grabowy C., Sager R. Differential activity of DNA methyltransferase in the life cycle of Chlamydomonas reinhardi. Proc Natl Acad Sci U S A. 1981 May;78(5):3118–3122. doi: 10.1073/pnas.78.5.3118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sano H., Noguchi H., Sager R. Characterization of DNA methyltransferase from bovine thymus cells. Eur J Biochem. 1983 Sep 15;135(2):181–185. doi: 10.1111/j.1432-1033.1983.tb07635.x. [DOI] [PubMed] [Google Scholar]
  23. Sano H., Royer H. D., Sager R. Identification of 5-methylcytosine in DNA fragments immobilized on nitrocellulose paper. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3581–3585. doi: 10.1073/pnas.77.6.3581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stewart C. L., Stuhlmann H., Jähner D., Jaenisch R. De novo methylation, expression, and infectivity of retroviral genomes introduced into embryonal carcinoma cells. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4098–4102. doi: 10.1073/pnas.79.13.4098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Swinton D. C., Hanawalt P. C. In vivo specific labeling of Chlamydomonas chloroplast DNA. J Cell Biol. 1972 Sep;54(3):592–597. doi: 10.1083/jcb.54.3.592. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES