title: Structural characterization of Set1 RNA recognition motifs and their role in histone H3 lysine 4 methylation.
abstract: The yeast Set1 histone H3 lysine 4 (H3K4) methyltransferase contains, in addition to its catalytic SET domain, a conserved RNA recognition motif (RRM1). We present here the crystal structure and the secondary structure assignment in solution of the Set1 RRM1. Although RRM1 has the expected betaalphabetabetaalphabeta RRM-fold, it lacks the typical RNA-binding features of these modules. RRM1 is not able to bind RNA by itself in vitro, but a construct combining RRM1 with a newly identified downstream RRM2 specifically binds RNA. In vivo, H3K4 methylation is not affected by a point mutation in RRM2 that preserves Set1 stability but affects RNA binding in vitro. In contrast mutating RRM1 destabilizes Set1 and leads to an increase of dimethylation of H3K4 at the 5'-coding region of active genes at the expense of trimethylation, whereas both, dimethylation decreases at the 3'-coding region. Taken together, our results suggest that Set1 RRMs bind RNA, but Set1 RNA-binding activity is not linked to H3K4 methylation.
authors.affiliation: Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, UMR8619 CNRS, Université de Paris-Sud, Orsay, France.
authors.forename: Lionel
authors.initials: L
authors.lastname: Trésaugues
authors.forename: Pierre-Marie
authors.initials: PM
authors.lastname: Dehé
authors.forename: Raphaël
authors.initials: R
authors.lastname: Guérois
authors.forename: Alfonso
authors.initials: A
authors.lastname: Rodriguez-Gil
authors.forename: Isabelle
authors.initials: I
authors.lastname: Varlet
authors.forename: Philippe
authors.initials: P
authors.lastname: Salah
authors.forename: Mercè
authors.initials: M
authors.lastname: Pamblanco
authors.forename: Pierre
authors.initials: P
authors.lastname: Luciano
authors.forename: Sophie
authors.initials: S
authors.lastname: Quevillon-Cheruel
authors.forename: Julie
authors.initials: J
authors.lastname: Sollier
authors.forename: Nicolas
authors.initials: N
authors.lastname: Leulliot
authors.forename: Joël
authors.initials: J
authors.lastname: Couprie
authors.forename: Vicente
authors.initials: V
authors.lastname: Tordera
authors.forename: Sophie
authors.initials: S
authors.lastname: Zinn-Justin
authors.forename: Sebastiàn
authors.initials: S
authors.lastname: Chàvez
authors.forename: Herman
authors.initials: H
authors.lastname: van Tilbeurgh
authors.forename: Vincent
authors.initials: V
authors.lastname: Géli
mesh_terms: D000595:Amino Acid Sequence; D017124:Conserved Sequence / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D011495:Histone-Lysine N-Methyltransferase; D006657:Histones / Q000737:chemistry* / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism*; D008745:Methylation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D021122:Protein Subunits / Q000378:metabolism; D012331:RNA, Fungal / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism*; D016415:Sequence Alignment; D013329:Structure-Activity Relationship; D013499:Surface Properties; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D006657:Histones; D021122:Protein Subunits; D012331:RNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D011495:Histone-Lysine N-Methyltransferase; C411250:SET1 protein, S cerevisiae; D008239:Lysine