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The spliceosomal proteins PPIH and PRPF4 exhibit bi-partite binding.
Rajiv, Caroline; Jackson, S RaElle; Cocklin, Simon; Eisenmesser, Elan Z; Davis, Tara L.
Afiliación
  • Rajiv C; Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, PA 19102, U.S.A.
  • Jackson SR; Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, PA 19102, U.S.A.
  • Cocklin S; Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, PA 19102, U.S.A.
  • Eisenmesser EZ; Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, U.S.A.
  • Davis TL; Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, PA 19102, U.S.A. tara.davis@drexelmed.edu.
Biochem J ; 474(21): 3689-3704, 2017 10 25.
Article en En | MEDLINE | ID: mdl-28935721
Pre-mRNA splicing is a dynamic, multistep process that is catalyzed by the RNA (ribonucleic acid)-protein complex called the spliceosome. The spliceosome contains a core set of RNAs and proteins that are conserved in all organisms that perform splicing. In higher organisms, peptidyl-prolyl isomerase H (PPIH) directly interacts with the core protein pre-mRNA processing factor 4 (PRPF4) and both integrate into the pre-catalytic spliceosome as part of the tri-snRNP (small nuclear RNA-protein complex) subcomplex. As a first step to understand the protein interactions that dictate PPIH and PRPF4 function, we expressed and purified soluble forms of each protein and formed a complex between them. We found two sites of interaction between PPIH and the N-terminus of PRPF4, an unexpected result. The N-terminus of PRPF4 is an intrinsically disordered region and does not adopt secondary structure in the presence of PPIH. In the absence of an atomic resolution structure, we used mutational analysis to identify point mutations that uncouple these two binding sites and find that mutations in both sites are necessary to break up the complex. A discussion of how this bipartite interaction between PPIH and PRPF4 may modulate spliceosomal function is included.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Empalme del ARN / Empalmosomas / Ribonucleoproteína Nuclear Pequeña U4-U6 / Isomerasa de Peptidilprolil Tipo de estudio: Prognostic_studies Idioma: En Revista: Biochem J Año: 2017 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Empalme del ARN / Empalmosomas / Ribonucleoproteína Nuclear Pequeña U4-U6 / Isomerasa de Peptidilprolil Tipo de estudio: Prognostic_studies Idioma: En Revista: Biochem J Año: 2017 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido