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Amino Acid Motifs
Cleavage And Polyadenylation Specificity Factor/*metabolism
Gene Expression Profiling
HEK293 Cells
HeLa Cells
Humans
Nuclear Proteins/*metabolism
Polyadenylation
Protein Binding
Protein Structure, Tertiary
RNA 3' End Processing/*physiology
RNA, Messenger/*metabolism
cleavage and polyadenylation
mRNA 3' processing
polyadenylation signal
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.
