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PMID:12556496

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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

Significance

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


See also

References

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