An interaction between U2AF 65 and CF I(m) links the splicing and 3' end processing machineries.
The protein factor U2 snRNP Auxiliary Factor (U2AF) 65 is an essential component required for splicing and involved in the coupling of splicing and 3' end processing of vertebrate pre-mRNAs. Here we have addressed the mechanisms by which U2AF 65 stimulates pre-mRNA 3' end processing. We identify an arginine/serine-rich region of U2AF 65 that mediates an interaction with an RS-like alternating charge domain of the 59 kDa subunit of the human cleavage factor I (CF I(m)), an essential 3' processing factor that functions at an early step in the recognition of the 3' end processing signal. Tethered functional analysis shows that the U2AF 65/CF I(m) 59 interaction stimulates in vitro 3' end cleavage and polyadenylation. These results therefore uncover a direct role of the U2AF 65/CF I(m) 59 interaction in the functional coordination of splicing and 3' end processing.
Millevoi
Stefania
S
INSERM U563, Toulouse, France.
Loulergue
Clarisse
C
Dettwiler
Sabine
S
Karaa
Sarah Zeïneb
SZ
Keller
Walter
W
Antoniou
Michael
M
Vagner
Stéphan
S
D020413:3' Untranslated Regions / Q000235:genetics / Q000378:metabolism*; D002474:Cell-Free System / Q000378:metabolism; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D026723:Polyadenylation / Q000502:physiology*; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D012326:RNA Splicing / Q000502:physiology*; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism*; D000072668:Splicing Factor U2AF; D020798:Two-Hybrid System Techniques; D039221:mRNA Cleavage and Polyadenylation Factors / Q000235:genetics / Q000378:metabolism*
D020413:3' Untranslated Regions; D009687:Nuclear Proteins; D012261:Ribonucleoproteins; D000072668:Splicing Factor U2AF; C000607494:U2AF2 protein, human; C450562:cleavage factor Im, human; D039221:mRNA Cleavage and Polyadenylation Factors