title: A complex secondary structure in U1A pre-mRNA that binds two molecules of U1A protein is required for regulation of polyadenylation.
abstract: The human U1A protein-U1A pre-mRNA complex and the relationship between its structure and function in inhibition of polyadenylation in vitro were investigated. Two molecules of U1A protein were shown to bind to a conserved region in the 3' untranslated region of U1A pre-mRNA. The secondary structure of this region was determined by a combination of theoretical prediction, phylogenetic sequence alignment, enzymatic structure probing and molecular genetics. The U1A binding sites form (part of) a complex secondary structure which is significantly different from the binding site of U1A protein on U1 snRNA. Studies with mutant pre-mRNAs showed that the integrity of much of this structure is required for both high affinity binding to U1A protein and specific inhibition of polyadenylation in vitro. In particular, binding of a single molecule of U1A protein to U1A pre-mRNA is not sufficient to produce efficient inhibition of polyadenylation.
authors.affiliation: University of Nijmegen, Department of Biochemistry, The Netherlands.
authors.forename: C W
authors.initials: CW
authors.lastname: van Gelder
authors.forename: S I
authors.initials: SI
authors.lastname: Gunderson
authors.forename: E J
authors.initials: EJ
authors.lastname: Jansen
authors.forename: W C
authors.initials: WC
authors.lastname: Boelens
authors.forename: M
authors.initials: M
authors.lastname: Polycarpou-Schwarz
authors.forename: I W
authors.initials: IW
authors.lastname: Mattaj
authors.forename: W J
authors.initials: WJ
authors.lastname: van Venrooij
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009694:Nucleic Acid Precursors / Q000378:metabolism; D011061:Poly A / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000378:metabolism; D016601:RNA-Binding Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
chemical_list: D009694:Nucleic Acid Precursors; D012333:RNA, Messenger; D012342:RNA, Small Nuclear; D016601:RNA-Binding Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear; C075339:U1A protein; D011061:Poly A