Relationships Between RNA Polymerase II Activity and Spt Elongation Factors to Spt- Phenotype and Growth in Saccharomyces cerevisiae.
The interplay between adjacent transcription units can result in transcription-dependent alterations in chromatin structure or recruitment of factors that determine transcription outcomes, including the generation of intragenic or other cryptic transcripts derived from cryptic promoters. Mutations in a number of genes in Saccharomyces cerevisiae confer both cryptic intragenic transcription and the Suppressor of Ty (Spt(-)) phenotype for the lys2-128∂ allele of the LYS2 gene. Mutants that suppress lys2-128∂ allow transcription from a normally inactive Ty1 ∂ promoter, conferring a LYS(+) phenotype. The arrangement of transcription units at lys2-128∂ is reminiscent of genes containing cryptic promoters within their open reading frames. We set out to examine the relationship between RNA Polymerase II (Pol II) activity, functions of Spt elongation factors, and cryptic transcription because of the previous observation that increased-activity Pol II alleles confer an Spt(-) phenotype. We identify both cooperating and antagonistic genetic interactions between Pol II alleles and alleles of elongation factors SPT4, SPT5, and SPT6 We find that cryptic transcription at FLO8 and STE11 is distinct from that at lys2-128∂, though all show sensitivity to reduction in Pol II activity, especially the expression of lys2-128∂ found in Spt(-) mutants. We determine that the lys2-128∂ Spt(-) phenotypes for spt6-1004 and increased activity rpo21/rpb1 alleles each require transcription from the LYS2 promoter. Furthermore, we identify the Ty1 transcription start site (TSS) within the ∂ element as the position of Spt(-) transcription in tested Spt(-) mutants.
Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843.
Ping
P
Cui
Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843.
Huiyan
H
Jin
Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843.
Manjula Ramya
MR
Vutukuru
Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843 cdkaplan@tamu.edu.
Craig D
0000-0002-7518-695X
CD
Kaplan
D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D056488:Histone Chaperones / Q000235:genetics / Q000378:metabolism; D020930:MAP Kinase Kinase Kinases / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D018626:Retroelements; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D024363:Transcription Initiation Site; D035602:Transcriptional Elongation Factors / Q000235:genetics / Q000378:metabolism*
D002868:Chromosomal Proteins, Non-Histone; C100940:FLO8 protein, S cerevisaie; D056488:Histone Chaperones; D009687:Nuclear Proteins; D018626:Retroelements; C080991:SPT4 protein, S cerevisiae; C065456:SPT6 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D035602:Transcriptional Elongation Factors; C069940:SPT5 transcriptional elongation factor; D020930:MAP Kinase Kinase Kinases; C067018:Ste11 protein, S cerevisiae; D012319:RNA Polymerase II; C511736:RPB1 protein, S cerevisiae
Ty1 element; cryptic transcription; gene expression; transcription elongation; transcription initiation