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PSF and p54nrb bind a conserved stem in U5 snRNA

Published online by Cambridge University Press:  03 October 2002

RUI PENG ,
BILLY T. DYE ,
ISMAEL PÉREZ ,
DARON C. BARNARD ,
AMANDA B. THOMPSON  and
JAMES G. PATTON
RUI PENG
Affiliation:
Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee 37235, USA
BILLY T. DYE
Affiliation:
Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee 37235, USA Present address: Institute for Molecular Virology, University of Wisconsin–Madison, Madison, Wisconsin 53706, USA
ISMAEL PÉREZ
Affiliation:
Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee 37235, USA
DARON C. BARNARD
Affiliation:
Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee 37235, USA Present address: Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA
AMANDA B. THOMPSON
Affiliation:
Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee 37235, USA
JAMES G. PATTON
Affiliation:
Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee 37235, USA
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Abstract

PTB-associated splicing factor (PSF) has been implicated in both early and late steps of pre-mRNA splicing, but its exact role in this process remains unclear. Here we show that PSF interacts with p54nrb, a highly related protein first identified based on cross-reactivity to antibodies against the yeast second-step splicing factor Prp18. We performed RNA-binding experiments to determine the preferred RNA-binding sequences for PSF and p54nrb, both individually and in combination. In all cases, iterative selection assays identified a purine-rich sequence located on the 3′ side of U5 snRNA stem 1b. Filter-binding assays and RNA affinity selection experiments demonstrated that PSF and p54nrb bind U5 snRNA with both the sequence and structure of stem 1b contributing to binding specificity. Sedimentation analyses show that both proteins associate with spliceosomes and with U4/U6.U5 tri-snPNP.

Keywords

p54nrbPSFsplicingU5 snRNA
Type
Research Article
Information
RNA , Volume 8 , Issue 10 , October 2002 , pp. 1334 - 1347
Copyright
2002 RNA Society

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