title: SET3p monomethylates histone H3 on lysine 9 and is required for the silencing of tandemly repeated transgenes in Chlamydomonas.
abstract: SET domain-containing proteins of the SU(VAR)3-9 class are major regulators of heterochromatin in several eukaryotes, including mammals, insects, plants and fungi. The function of these polypeptides is mediated, at least in part, by their ability to methylate histone H3 on lysine 9 (H3K9). Indeed, mutants defective in SU(VAR)3-9 proteins have implicated di- and/or trimethyl H3K9 in the formation and/or maintenance of heterochromatin across the eukaryotic spectrum. Yet, the biological significance of monomethyl H3K9 has remained unclear because of the lack of mutants exclusively defective in this modification. Interestingly, a SU(VAR)3-9 homolog in the unicellular green alga Chlamydomonas reinhardtii, SET3p, functions in vitro as a specific H3K9 monomethyltransferase. RNAi-mediated suppression of SET3 reactivated the expression of repetitive transgenic arrays and reduced global monomethyl H3K9 levels. Moreover, chromatin immunoprecipitation (ChIP) assays demonstrated that transgene reactivation correlated with the partial loss of monomethyl H3K9 from their chromatin. In contrast, the levels of trimethyl H3K9 or the repression of euchromatic sequences were not affected by SET3 downregulation; whereas dimethyl H3K9 was undetectable in Chlamydomonas. Thus, our observations are consistent with a role for monomethyl H3K9 as an epigenetic mark of repressed chromatin and raise questions as to the functional distinctiveness of different H3K9 methylation states.
authors.affiliation: School of Biological Sciences and Plant Science Initiative, University of Nebraska, Lincoln, NE 68588-0666, USA.
authors.forename: J Armando
authors.initials: JA
authors.lastname: Casas-Mollano
authors.forename: Karin
authors.initials: K
authors.lastname: van Dijk
authors.forename: John
authors.initials: J
authors.lastname: Eisenhart
authors.forename: Heriberto
authors.initials: H
authors.lastname: Cerutti
mesh_terms: D020418:Algal Proteins / Q000145:classification / Q000235:genetics / Q000502:physiology*; D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000201:enzymology* / Q000235:genetics*; D002843:Chromatin / Q000378:metabolism; D018899:CpG Islands; D019175:DNA Methylation; D018628:Gene Dosage; D020868:Gene Silencing; D011495:Histone-Lysine N-Methyltransferase / Q000145:classification / Q000235:genetics / Q000502:physiology*; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000145:classification; D034622:RNA Interference; D017386:Sequence Homology, Amino Acid; D020080:Tandem Repeat Sequences; D019076:Transgenes
chemical_list: D020418:Algal Proteins; D002843:Chromatin; D006657:Histones; D010940:Plant Proteins; D011495:Histone-Lysine N-Methyltransferase; D008239:Lysine