CTD serine-2 plays a critical role in splicing and termination factor recruitment to RNA polymerase II in vivo.
    
    
        
    
    
        
        Nucleic Acids Res. 41, 1591-1603 (2013)
    
    
    
      
      
	
	    Co-transcriptional pre-mRNA processing relies on reversible phosphorylation of the carboxyl-terminal domain (CTD) of Rpb1, the largest subunit of RNA polymerase II (RNAP II). In this study, we replaced in live cells the endogenous Rpb1 by S2A Rpb1, where the second serines (Ser2) in the CTD heptapeptide repeats were switched to alanines, to prevent phosphorylation. Although slower, S2A RNAP II was able to transcribe. However, it failed to recruit splicing components such as U2AF65 and U2 snRNA to transcription sites, although the recruitment of U1 snRNA was not affected. As a consequence, co-transcriptional splicing was impaired. Interestingly, the magnitude of the S2A RNAP II splicing defect was promoter dependent. In addition, S2A RNAP II showed an impaired recruitment of the cleavage factor PCF11 to pre-mRNA and a defect in 3'-end RNA cleavage. These results suggest that CTD Ser2 plays critical roles in co-transcriptional pre-mRNA maturation in vivo: It likely recruits U2AF65 to ensure an efficient co-transcriptional splicing and facilitates the recruitment of pre-mRNA 3'-end processing factors to enhance 3'-end cleavage.
	
	
	    
	
       
      
	
	    
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        Publication type
        Article: Journal article
    
 
    
        Document type
        Scientific Article
    
 
    
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        Keywords
        Pre-messenger-rna ; P-tefb Kinase ; 3' End ; Transcription Elongation ; Gene-expression ; Fission Yeast ; Domain ; Phosphorylation ; Complex ; Recognition
    
 
    
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        Language
        english
    
 
    
        Publication Year
        2013
    
 
    
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        HGF-reported in Year
        2013
    
 
    
    
        ISSN (print) / ISBN
        0305-1048
    
 
    
        e-ISSN
        1362-4962
    
 
    
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	    Volume: 41,  
	    Issue: 3,  
	    Pages: 1591-1603 
	    Article Number: ,  
	    Supplement: ,  
	
    
 
    
        
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            Oxford University Press
        
 
        
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        Reviewing status
        Peer reviewed
    
 
     
    
        POF-Topic(s)
        30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
    
 
    
        Research field(s)
        Immune Response and Infection
    
 
    
        PSP Element(s)
        G-501490-001
    
 
    
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        Erfassungsdatum
        2013-04-11