HnRNP A1 Proofreads 3' Splice Site Recognition by U2AF

Joao Paulo Tavanez, Tobias Madl, Hamed Kooshapur, Michael Sattler, Juan Valcárcel

Research output: Contribution to journalArticlepeer-review

79 Scopus citations

Abstract

One of the earliest steps in metazoan pre-mRNA splicing involves binding of U2 snRNP auxiliary factor (U2AF) 65 KDa subunit to the polypyrimidine (Py) tract and of the 35 KDa subunit to the invariant AG dinucleotide at the intron 3' end. Here we use invitro and invivo depletion, as well as reconstitution assays using purified components, to identify hnRNP A1 as an RNA binding protein that allows U2AF todiscriminate between pyrimidine-rich RNA sequences followed or not by a 3' splice site AG. Biochemical and NMR data indicate that hnRNP A1 forms a ternary complex with the U2AF heterodimer on AG-containing/uridine-rich RNAs, while it displaces U2AF from non-AG-containing/uridine-rich RNAs, an activity that requires the glycine-rich domain of hnRNP A1. Consistent with the functional relevance of this activity for splicing, proofreading assays reveal a role for hnRNP A1 in U2AF-mediated recruitment of U2 snRNP to the pre-mRNA.

Original languageEnglish
Pages (from-to)314-329
Number of pages16
JournalMolecular Cell
Volume45
Issue number3
DOIs
StatePublished - 10 Feb 2012

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