A pathway of sequential arginine-serine-rich domain-splicing signal interactions during mammalian spliceosome assembly

UMMS Affiliation

Program in Molecular Medicine; Program in Gene Function and Expression

Publication Date


Document Type



Arginine; Cross-Linking Reagents; Enhancer Elements, Genetic; Exons; Hela Cells; Humans; Nuclear Proteins; Plasmids; Protein Structure, Tertiary; *RNA Precursors; RNA Splicing; RNA, Messenger; Ribonucleoproteins; Serine; Signal Transduction; Spliceosomes; Transcription, Genetic


Genetics and Genomics


Serine-arginine (SR) proteins are general splicing factors and can function through binding to exonic splicing enhancers (ESEs). SR proteins and several other mammalian splicing factors contain an arginine-serine-rich (RS) domain required to promote splicing. We have recently found that the ESE bound RS domain functions by contacting the branchpoint. Here, we perform RNA-protein crosslinking experiments to show that the branchpoint is sequentially contacted first in complex E by the RS domain of the essential splicing factor U2AF(65) and then in the prespliceosome by the ESE bound RS domain. Although the ESE bound RS domain can promote formation of the prespliceosome, at least one additional SR protein is required for complete spliceosome assembly. We show that the RS domain of this additional SR protein contacts the 5' splice site specifically in the mature spliceosome. We propose that direct contact with splicing signals is a general mechanism by which RS domains promote splicing.

DOI of Published Version



Mol Cell. 2004 Nov 5;16(3):363-73. Link to article on publisher's site

Journal/Book/Conference Title

Molecular cell

Related Resources

Link to Article in PubMed

PubMed ID