The alternative splicing of human genes is dependent on SR proteins, a family of essential splicing factors whose name derives from a signature C-terminal domain rich in arginine–serine dipeptide repeats (RS domains). Although the SRPKs (SR-specific protein kinases) phosphorylate these repeats, RS domains also contain prolines with flanking serines that are phosphorylated by a second family of protein kinases known as the CLKs (Cdc2-like kinases). The role of specific serine–proline phosphorylation within the RS domain has been difficult to assign since CLKs also phosphorylate arginine–serine dipeptides and, thus, display overlapping residue specificities with the SRPKs. In the present study, we address the effects of discrete serine–proline phosphorylation on the conformation and cellular function of the SR protein SRSF1 (SR protein splicing factor 1). Using chemical tagging and dephosphorylation experiments, we show that modification of serine–proline dipeptides broadly amplifies the conformational ensemble of SRSF1. The induction of these new structural forms triggers SRSF1 mobilization in the nucleus and alters its binding mechanism to an exonic splicing enhancer in precursor mRNA. These physical events correlate with changes in the alternative splicing of over 100 human genes based on a global splicing assay. Overall, these studies draw a direct causal relationship between a specific type of chemical modification in an SR protein and the regulation of alternative gene splicing programmes.
Skip Nav Destination
Article navigation
March 2015
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
February 20 2015
Conserved proline-directed phosphorylation regulates SR protein conformation and splicing function
Malik M. Keshwani;
Malik M. Keshwani
*Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093-0636, U.S.A.
Search for other works by this author on:
Brandon E. Aubol;
Brandon E. Aubol
*Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093-0636, U.S.A.
Search for other works by this author on:
Laurent Fattet;
Laurent Fattet
*Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093-0636, U.S.A.
Search for other works by this author on:
Chen-Ting Ma;
Chen-Ting Ma
*Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093-0636, U.S.A.
Search for other works by this author on:
Jinsong Qiu;
Jinsong Qiu
†Department of Cellular & Molecular Medicine, University of California, San Diego, La Jolla, CA 92093-0636, U.S.A.
Search for other works by this author on:
Patricia A. Jennings;
Patricia A. Jennings
‡Department of Chemistry & Biochemistry, University of California, San Diego, La Jolla, CA 92093-0636, U.S.A.
Search for other works by this author on:
Xiang-Dong Fu;
Xiang-Dong Fu
†Department of Cellular & Molecular Medicine, University of California, San Diego, La Jolla, CA 92093-0636, U.S.A.
Search for other works by this author on:
Joseph A. Adams
Joseph A. Adams
1
*Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093-0636, U.S.A.
1To whom correspondence should be addressed (email: j2adams@ucsd.edu.).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
November 05 2014
Revision Received:
December 17 2014
Accepted:
December 22 2014
Accepted Manuscript online:
December 22 2014
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2015 Biochemical Society
2015
Biochem J (2015) 466 (2): 311–322.
Article history
Received:
November 05 2014
Revision Received:
December 17 2014
Accepted:
December 22 2014
Accepted Manuscript online:
December 22 2014
Citation
Malik M. Keshwani, Brandon E. Aubol, Laurent Fattet, Chen-Ting Ma, Jinsong Qiu, Patricia A. Jennings, Xiang-Dong Fu, Joseph A. Adams; Conserved proline-directed phosphorylation regulates SR protein conformation and splicing function. Biochem J 1 March 2015; 466 (2): 311–322. doi: https://doi.org/10.1042/BJ20141373
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.