Mutations in RRM4 uncouple the splicing repression and RNA-binding activities of polypyrimidine tract binding protein

Haiying Liu, Wenqing Zhang, Robyn Reed, Weiqun Liu, Paula J. Grabowski

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

The polypyrimidine tract binding protein (PTB, or hnRNP I) contains four RNA-binding domains of the ribonucleoprotein fold type (RRMs 1,2,3, and 4), and mediates the negative regulation of alternative splicing through sequence-specific binding to intronic splicing repressor elements. To assess the roles of individual RRM domains in splicing repression, a neural-specific splicing extract was used to screen for loss-of-function mutations that fail to switch splicing from the neural to nonneural pathway. These results show that three RRMs are sufficient for wild-type RNA binding and splicing repression activity, provided that RRM4 is intact. Surprisingly, the deletion of RRM4, or as few as 12 RRM4 residues, effectively uncouples these functions. Such an uncoupling phenotype is unique to RRM4, and suggests a possible regulatory role for this domain either in mediating specific RNA contacts, and/or contacts with putative splicing corepressors. Evidence of a role for RRM4 in anchoring PTB binding adjacent to the branch site is shown by mobility shift and RNA footprinting assays.

Original languageEnglish (US)
Pages (from-to)137-149
Number of pages13
JournalRNA
Volume8
Issue number2
DOIs
StatePublished - Apr 18 2002

Fingerprint

Polypyrimidine Tract-Binding Protein
RNA Splicing
RNA
Heterogeneous-Nuclear Ribonucleoproteins
Co-Repressor Proteins
Mutation
Ribonucleoproteins
Alternative Splicing
Phenotype

All Science Journal Classification (ASJC) codes

  • Molecular Biology

Cite this

Liu, Haiying ; Zhang, Wenqing ; Reed, Robyn ; Liu, Weiqun ; Grabowski, Paula J. / Mutations in RRM4 uncouple the splicing repression and RNA-binding activities of polypyrimidine tract binding protein. In: RNA. 2002 ; Vol. 8, No. 2. pp. 137-149.
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Mutations in RRM4 uncouple the splicing repression and RNA-binding activities of polypyrimidine tract binding protein. / Liu, Haiying; Zhang, Wenqing; Reed, Robyn; Liu, Weiqun; Grabowski, Paula J.

In: RNA, Vol. 8, No. 2, 18.04.2002, p. 137-149.

Research output: Contribution to journalArticle

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