Rad6 plays a role in transcriptional activation through ubiquitylation of histone H2B.
Covalent modifications of the histone N tails play important roles in eukaryotic gene expression. Histone acetylation, in particular, is required for the activation of a subset of eukaryotic genes through the targeted recruitment of histone acetyltransferases. We have reported that a histone C tail modification, ubiquitylation of H2B, is required for optimal expression of several inducible yeast genes, consistent with a role in transcriptional activation. H2B was shown to be ubiquitylated and then deubiquitylated at the GAL1 core promoter following galactose induction. We now show that the Rad6 protein, which catalyzes monoubiquitylation of H2B, is transiently associated with the GAL1 promoter upon gene activation, and that the period of its association temporally overlaps with the period of H2B ubiquitylation. Rad6 promoter association depends on the Gal4 activator and the Rad6-associated E3 ligase, Bre1, but is independent of the histone acetyltransferase, Gcn5. The SAGA complex, which contains a ubiquitin protease that targets H2B for deubiquitylation, is recruited to the GAL1 promoter in the absence of H2B ubiquitylation. The data suggest that Rad6 and SAGA function independently during galactose induction, and that the staged recruitment of these two factors to the GAL1 promoter regulates the ubiquitylation and deubiquitylation of H2B. We additionally show that both Rad6 and ubiquitylated H2B are absent from two regions of transcriptionally silent chromatin but present at genes that are actively transcribed. Thus, like histone H3 lysine 4 and lysine 79 methylation, two modifications that it regulates, Rad6-directed H2B ubiquitylation defines regions of active chromatin.
Kao
Cheng-Fu
CF
University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA.
Hillyer
Cory
C
Tsukuda
Toyoko
T
Henry
Karl
K
Berger
Shelley
S
Osley
Mary Ann
MA
D000135:Acid Phosphatase / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D006657:Histones / Q000378:metabolism*; D046911:Macromolecular Substances; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D015533:Transcriptional Activation / Q000502:physiology*; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism*; D014452:Ubiquitins / Q000378:metabolism*
D006657:Histones; D046911:Macromolecular Substances; D029701:Saccharomyces cerevisiae Proteins; D014452:Ubiquitins; C471606:RAD6 protein, S cerevisiae; D044763:Ubiquitin-Conjugating Enzymes; D000135:Acid Phosphatase