The RAD6/BRE1 histone modification pathway in Saccharomyces confers radiation resistance through a RAD51-dependent process that is independent of RAD18.
We examine ionizing radiation (IR) sensitivity and epistasis relationships of several Saccharomyces mutants affecting post-translational modifications of histones H2B and H3. Mutants bre1Delta, lge1Delta, and rtf1Delta, defective in histone H2B lysine 123 ubiquitination, show IR sensitivity equivalent to that of the dot1Delta mutant that we reported on earlier, consistent with published findings that Dot1p requires H2B K123 ubiquitination to fully methylate histone H3 K79. This implicates progressive K79 methylation rather than mono-methylation in IR resistance. The set2Delta mutant, defective in H3 K36 methylation, shows mild IR sensitivity whereas mutants that abolish H3 K4 methylation resemble wild type. The dot1Delta, bre1Delta, and lge1Delta mutants show epistasis for IR sensitivity. The paf1Delta mutant, also reportedly defective in H2B K123 ubiquitination, confers no sensitivity. The rad6Delta, rad51null, rad50Delta, and rad9Delta mutations are epistatic to bre1Delta and dot1Delta, but rad18Delta and rad5Delta show additivity with bre1Delta, dot1Delta, and each other. The bre1Delta rad18Delta double mutant resembles rad6Delta in sensitivity; thus the role of Rad6p in ubiquitinating H2B accounts for its extra sensitivity compared to rad18Delta. We conclude that IR resistance conferred by BRE1 and DOT1 is mediated through homologous recombinational repair, not postreplication repair, and confirm findings of a G1 checkpoint role for the RAD6/BRE1/DOT1 pathway.
Game
John C
JC
Life Sciences Division, Lawrence Berkeley National Laboratory, CA 94720, USA. jcgame@lbl.gov
Williamson
Marsha S
MS
Spicakova
Tatiana
T
Brown
J Martin
JM
D004260:DNA Repair / Q000235:genetics* / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D016193:G1 Phase / Q000187:drug effects / Q000235:genetics; D017353:Gene Deletion; D006657:Histones / Q000235:genetics / Q000378:metabolism; D008745:Methylation / Q000528:radiation effects; D011499:Protein Processing, Post-Translational / Q000235:genetics; D051135:Rad51 Recombinase / Q000235:genetics* / Q000378:metabolism; D011839:Radiation, Ionizing; D011995:Recombination, Genetic / Q000235:genetics / Q000528:radiation effects; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics* / Q000378:metabolism
C472168:Bre1 protein, S cerevisiae; D004268:DNA-Binding Proteins; D006657:Histones; C063607:RAD18 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C471606:RAD6 protein, S cerevisiae; D044763:Ubiquitin-Conjugating Enzymes; C495151:RAD51 protein, S cerevisiae; D051135:Rad51 Recombinase