title: Poly(A) tail length is controlled by the nuclear poly(A)-binding protein regulating the interaction between poly(A) polymerase and the cleavage and polyadenylation specificity factor.
abstract: Poly(A) tails of mRNAs are synthesized in the cell nucleus with a defined length, approximately 250 nucleotides in mammalian cells. The same type of length control is seen in an in vitro polyadenylation system reconstituted from three proteins: poly(A) polymerase, cleavage and polyadenylation specificity factor (CPSF), and the nuclear poly(A)-binding protein (PABPN1). CPSF, binding the polyadenylation signal AAUAAA, and PABPN1, binding the growing poly(A) tail, cooperatively stimulate poly(A) polymerase such that a complete poly(A) tail is synthesized in one processive event, which terminates at a length of approximately 250 nucleotides. We report that PABPN1 is required to restrict CPSF binding to the AAUAAA sequence and to permit the stimulation of poly(A) polymerase by AAUAAA-bound CPSF to be maintained throughout the elongation reaction. The stimulation by CPSF is disrupted when the poly(A) tail has reached a length of approximately 250 nucleotides, and this terminates processive elongation. PABPN1 measures the length of the tail and is responsible for disrupting the CPSF-poly(A) polymerase interaction.
authors.affiliation: Institute of Biochemistry and Biotechnology, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Strasse 3, 06120 Halle, Germany.
authors.forename: Uwe
authors.initials: U
authors.lastname: Kühn
authors.forename: Miriam
authors.initials: M
authors.lastname: Gündel
authors.forename: Anne
authors.initials: A
authors.lastname: Knoth
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Kerwitz
authors.forename: Sabine
authors.initials: S
authors.lastname: Rüdel
authors.forename: Elmar
authors.initials: E
authors.lastname: Wahle
mesh_terms: D000818:Animals; D002417:Cattle; D039223:Cleavage And Polyadenylation Specificity Factor / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D011061:Poly A / Q000737:chemistry / Q000378:metabolism*; D039122:Poly(A)-Binding Protein I / Q000378:metabolism*; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism*; D011485:Protein Binding; D039104:RNA 3' Polyadenylation Signals / Q000235:genetics
chemical_list: D039223:Cleavage And Polyadenylation Specificity Factor; D039122:Poly(A)-Binding Protein I; D011061:Poly A; D011062:Polynucleotide Adenylyltransferase