PMID:12556496

Citation	Lindstrom, DL, Squazzo, SL, Muster, N, Burckin, TA, Wachter, KC, Emigh, CA, McCleery, JA, Yates, JR 3rd and Hartzog, GA (2003) Dual roles for Spt5 in pre-mRNA processing and transcription elongation revealed by identification of Spt5-associated proteins. Mol. Cell. Biol. 23:1368-78
Abstract	During transcription elongation, eukaryotic RNA polymerase II (Pol II) must contend with the barrier presented by nucleosomes. The conserved Spt4-Spt5 complex has been proposed to regulate elongation through nucleosomes by Pol II. To help define the mechanism of Spt5 function, we have characterized proteins that coimmunopurify with Spt5. Among these are the general elongation factors TFIIF and TFIIS as well as Spt6 and FACT, factors thought to regulate elongation through nucleosomes. Spt5 also coimmunopurified with the mRNA capping enzyme and cap methyltransferase, and spt4 and spt5 mutations displayed genetic interactions with mutations in capping enzyme genes. Additionally, we found that spt4 and spt5 mutations lead to accumulation of unspliced pre-mRNA. Spt5 also copurified with several previously unstudied proteins; we demonstrate that one of these is encoded by a new member of the SPT gene family. Finally, by immunoprecipitating these factors we found evidence that Spt5 participates in at least three Pol II complexes. These observations provide new evidence of roles for Spt4-Spt5 in pre-mRNA processing and transcription elongation.
Links	PubMed PMC141151
Keywords	Carrier Proteins/metabolism; Cell Cycle Proteins/metabolism; Chromosomal Proteins, Non-Histone; Methyltransferases/isolation & purification; Methyltransferases/metabolism; Mutation; Nuclear Proteins/genetics; Nuclear Proteins/metabolism; Nucleotidyltransferases/isolation & purification; Nucleotidyltransferases/metabolism; RNA Polymerase II/isolation & purification; RNA Polymerase II/metabolism; RNA Precursors; RNA Processing, Post-Transcriptional; RNA Splicing; RNA, Messenger/metabolism; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/metabolism; Transcription Factors/metabolism; Transcription Factors, General/isolation & purification; Transcription Factors, General/metabolism; Transcription Factors, TFII/isolation & purification; Transcription Factors, TFII/metabolism; Transcriptional Elongation Factors/genetics; Transcriptional Elongation Factors/isolation & purification; Transcriptional Elongation Factors/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
YEAST:SPT4	involved_in	GO:0000398: mRNA splicing, via spliceosome	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
YEAST:SPT4	involved_in	GO:0006370: 7-methylguanosine mRNA capping	ECO:0000316: genetic interaction evidence used in manual assertion	SGD:S000003098	P		Seeded From UniProt	complete
YEAST:IWS1	located_in	GO:0005634: nucleus	ECO:0000305: curator inference used in manual assertion	GO:0001085 GO:0006357	C		Seeded From UniProt	complete
YEAST:IWS1	enables	GO:0000993: RNA polymerase II complex binding	ECO:0000353: physical interaction evidence used in manual assertion	SGD:S000002299 SGD:S000005677	F		Seeded From UniProt	complete
YEAST:IWS1	enables	GO:0001085: RNA polymerase II transcription factor binding	ECO:0000353: physical interaction evidence used in manual assertion	SGD:S000004470	F		Seeded From UniProt	complete
YEAST:IWS1	involved_in	GO:0006357: regulation of transcription by RNA polymerase II	ECO:0000316: genetic interaction evidence used in manual assertion	SGD:S000003295 SGD:S000004470	P		Seeded From UniProt	complete
YEAST:IWS1	involved_in	GO:0006357: regulation of transcription by RNA polymerase II	ECO:0000353: physical interaction evidence used in manual assertion	SGD:S000002299 SGD:S000004470 SGD:S000005677	P		Seeded From UniProt	complete

References

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