title: The yeast transcription elongation factor Spt4/5 is a sequence-specific RNA binding protein.
abstract: The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co-transcriptional pre-mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence-specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA-binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5.
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, Crawley, Western Australia, 6009, Australia.
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Blythe
authors.affiliation: Department of Molecular, Cell and Developmental Biology, University of California, Santa Cruz, California, 95064.
authors.forename: Berra
authors.initials: B
authors.lastname: Yazar-Klosinski
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Webster
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Cruz, California, 95064.
authors.forename: Eefei
authors.initials: E
authors.lastname: Chen
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia.
authors.forename: Marylène
authors.initials: M
authors.lastname: Vandevenne
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia.
authors.forename: Katerina
authors.initials: K
authors.lastname: Bendak
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, New South Wales, 2006, Australia.
authors.forename: Joel P
authors.initials: JP
authors.lastname: Mackay
authors.affiliation: Department of Molecular, Cell and Developmental Biology, University of California, Santa Cruz, California, 95064.
authors.forename: Grant A
authors.initials: GA
authors.lastname: Hartzog
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, Crawley, Western Australia, 6009, Australia.
authors.forename: Alice
authors.initials: A
authors.lastname: Vrielink
mesh_terms: D000818:Animals; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D000072417:Protein Domains; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D035602:Transcriptional Elongation Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; C080991:SPT4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D035602:Transcriptional Elongation Factors; C069940:SPT5 transcriptional elongation factor; D012319:RNA Polymerase II
keywords: RNA binding; RNA polymerase; SELEX; Spt4/5; transcription elongation; transcription elongation factor