Activated transcription independent of the RNA polymerase II holoenzyme in budding yeast.
We investigated whether the multisubunit holoenzyme complex of RNA polymerase II (Pol II) and mediator is universally required for transcription in budding yeast. DeltaCTD Pol II lacking the carboxy-terminal domain of the large subunit cannot assemble with mediator but can still transcribe the CUP1 gene. CUP1 transcripts made by DeltaCTD Pol II initiated correctly and some extended past the normal poly(A) site yielding a novel dicistronic mRNA. Most CUP1 transcripts made by DeltaCTD Pol II were degraded but could be stabilized by deletion of the XRN1 gene. Unlike other genes, transcription of CUP1 and HSP82 also persisted after inactivation of the CTD kinase Kin28 or the mediator subunit Srb4. The upstream-activating sequence (UAS) of the CUP1 promoter was sufficient to drive Cu2+ inducible transcription without Srb4 and heat shock inducible transcription without the CTD. We conclude that the Pol II holoenzyme is not essential for all UAS-dependent activated transcription in yeast.
Amgen Institute, Ontario Cancer Institute, Toronto, Ontario M5G 2C1, Canada.
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McNeil
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Agah
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Bentley
D002352:Carrier Proteins; D003067:Coenzymes / Q000378:metabolism*; D018844:Cyclin-Dependent Kinases; D005656:Fungal Proteins / Q000502:physiology; D015966:Gene Expression Regulation, Fungal; D056892:Mediator Complex; D008668:Metallothionein / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000502:physiology; D012319:RNA Polymerase II / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000502:physiology; D015533:Transcriptional Activation
C500769:CUP1-1 protein, S cerevisiae; D002352:Carrier Proteins; D003067:Coenzymes; D005656:Fungal Proteins; D056892:Mediator Complex; D012333:RNA, Messenger; C082148:SRB4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C007121:copper thionein; D008668:Metallothionein; D017346:Protein Serine-Threonine Kinases; D018844:Cyclin-Dependent Kinases; C084210:Kin28 protein kinase, S cerevisiae; D012319:RNA Polymerase II