title: CPSF30 and Wdr33 directly bind to AAUAAA in mammalian mRNA 3' processing.
abstract: AAUAAA is the most highly conserved motif in eukaryotic mRNA polyadenylation sites and, in mammals, is specifically recognized by the multisubunit CPSF (cleavage and polyadenylation specificity factor) complex. Despite its critical functions in mRNA 3' end formation, the molecular basis for CPSF-AAUAAA interaction remains poorly defined. The CPSF subunit CPSF160 has been implicated in AAUAAA recognition, but direct evidence has been lacking. Using in vitro and in vivo assays, we unexpectedly found that CPSF subunits CPSF30 and Wdr33 directly contact AAUAAA. Importantly, the CPSF30-RNA interaction is essential for mRNA 3' processing and is primarily mediated by its zinc fingers 2 and 3, which are specifically targeted by the influenza protein NS1A to suppress host mRNA 3' processing. Our data suggest that AAUAAA recognition in mammalian mRNA 3' processing is more complex than previously thought and involves multiple protein-RNA interactions.
authors.affiliation: Department of Microbiology and Molecular Genetics, School of Medicine, University of California at Irvine, Irvine, California 92697, USA;
authors.forename: Serena L
authors.initials: SL
authors.lastname: Chan
authors.affiliation: Department of Molecular Neuroscience, University College London Institute of Neurology, London WC1N 3BG, United Kingdom; Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom;
authors.forename: Ina
authors.initials: I
authors.lastname: Huppertz
authors.affiliation: Department of Microbiology and Molecular Genetics, School of Medicine, University of California at Irvine, Irvine, California 92697, USA;
authors.forename: Chengguo
authors.initials: C
authors.lastname: Yao
authors.affiliation: Department of Microbiology and Molecular Genetics, School of Medicine, University of California at Irvine, Irvine, California 92697, USA; Institute for Genomics and Bioinformatics, Department of Computer Science, University of California at Irvine Irvine, California 92697, USA;
authors.forename: Lingjie
authors.initials: L
authors.lastname: Weng
authors.affiliation: Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA;
authors.forename: James J
authors.initials: JJ
authors.lastname: Moresco
authors.affiliation: Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA;
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
authors.affiliation: Department of Molecular Neuroscience, University College London Institute of Neurology, London WC1N 3BG, United Kingdom; Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom;
authors.forename: Jernej
authors.initials: J
authors.lastname: Ule
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York 10027, USA.
authors.forename: James L
authors.initials: JL
authors.lastname: Manley
authors.affiliation: Department of Microbiology and Molecular Genetics, School of Medicine, University of California at Irvine, Irvine, California 92697, USA;
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Shi
mesh_terms: D020816:Amino Acid Motifs; D039223:Cleavage And Polyadenylation Specificity Factor / Q000378:metabolism*; D020869:Gene Expression Profiling; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D026723:Polyadenylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D026721:RNA 3' End Processing / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism*
chemical_list: C000594014:CPSF4 protein, human; D039223:Cleavage And Polyadenylation Specificity Factor; D009687:Nuclear Proteins; D012333:RNA, Messenger; C486022:WDR33 protein, human
keywords: cleavage and polyadenylation; mRNA 3′ processing; polyadenylation signal