Mutations in RNA polymerase II and elongation factor SII severely reduce mRNA levels in Saccharomyces cerevisiae.
Elongation factor SII interacts with RNA polymerase II and enables it to transcribe through arrest sites in vitro. The set of genes dependent upon SII function in vivo and the effects on RNA levels of mutations in different components of the elongation machinery are poorly understood. Using yeast lacking SII and bearing a conditional allele of RPB2, the gene encoding the second largest subunit of RNA polymerase II, we describe a genetic interaction between SII and RPB2. An SII gene disruption or the rpb2-10 mutation, which yields an arrest-prone enzyme in vitro, confers sensitivity to 6-azauracil (6AU), a drug that depresses cellular nucleoside triphosphates. Cells with both mutations had reduced levels of total poly(A)+ RNA and specific mRNAs and displayed a synergistic level of drug hypersensitivity. In cells in which the SII gene was inactivated, rpb2-10 became dominant, as if template-associated mutant RNA polymerase II hindered the ability of wild-type polymerase to transcribe. Interestingly, while 6AU depressed RNA levels in both wild-type and mutant cells, wild-type cells reestablished normal RNA levels, whereas double-mutant cells could not. This work shows the importance of an optimally functioning elongation machinery for in vivo RNA synthesis and identifies an initial set of candidate genes with which SII-dependent transcription can be studied.
Lennon
J C
JC
Graduate Program in Genetics and Molecular Biology and Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Wind
M
M
Saunders
L
L
Hock
M B
MB
Reines
D
D
D002455:Cell Division / Q000187:drug effects; D009154:Mutation; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism; D012331:RNA, Fungal / Q000378:metabolism*; D012333:RNA, Messenger; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D035165:Transcription Factors, General; D035602:Transcriptional Elongation Factors; D014498:Uracil / Q000031:analogs & derivatives / Q000494:pharmacology
D012331:RNA, Fungal; D012333:RNA, Messenger; D014157:Transcription Factors; D035165:Transcription Factors, General; D035602:Transcriptional Elongation Factors; C018648:transcription factor S-II; D014498:Uracil; D012319:RNA Polymerase II; C100248:azauracil