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PMID: 21343338 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

RNA helicase p68 (DDX5) regulates tau exon 10 splicing by modulating a stem-loop structure at the 5' splice site.

Molecular and cellular biology ·Vol. 31 ·No. 9 ·2011-05-00 ·Pages 1812-21

Kar A, Fushimi K, Zhou X, Ray P, Shi C, Chen X, Liu Z, Chen S, Wu JY

Abstract

Regulation of tau exon 10 splicing plays an important role in tauopathy. One of the cis elements regulating tau alternative splicing is a stem-loop structure at the 5' splice site of tau exon 10. The RNA helicase(s) modulating this stem-loop structure was unknown. We searched for splicing regulators interacting with this stem-loop region using an RNA affinity pulldown-coupled mass spectrometry approach and identified DDX5/RNA helicase p68 as an activator of tau exon 10 splicing. The activity of p68 in stimulating tau exon 10 inclusion is dependent on RBM4, an intronic splicing activator. RNase H cleavage and U1 protection assays suggest that p68 promotes conformational change of the stem-loop structure, thereby increasing the access of U1snRNP to the 5' splice site of tau exon 10. This study reports the first RNA helicase interacting with a stem-loop structure at the splice site and regulating alternative splicing in a helicase-dependent manner. Our work uncovers a previously unknown function of p68 in regulating tau exon 10 splicing. Furthermore, our experiments reveal functional interaction between two splicing activators for tau exon 10, p68 binding at the stem-loop region and RBM4 interacting with the intronic splicing enhancer region.

MeSH Terms
Cell Line DEAD-box RNA Helicases/metabolism Exons Humans Nucleic Acid Conformation RNA Splicing RNA-Binding Proteins/metabolism Ribonucleoproteins, Small Nuclear/metabolism tau Proteins/genetics
Chemicals
RBM4 protein, human RNA-Binding Proteins Ribonucleoproteins, Small Nuclear SNRNP35 protein, human tau Proteins Ddx5 protein, human DEAD-box RNA Helicases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Kar Amar
Northwestern University Feinberg School of Medicine, 303 E. Superior St., Lurie 6-117, Chicago, IL 60611, USA.
Fushimi Kazuo
Zhou Xiaohong
Ray Payal
Shi Chen
Chen Xiaoping
Liu Zhiren
Chen She
Wu Jane Y
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01149-10
Published
2011-05-00
Epub
2011-00-22
Pages
1812-21
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC3133221
Subset
IM
Grants
NIA NIH HHS · R01 AG017518 · United States
NIA NIH HHS · R01 AG033004 · United States
NCI NIH HHS · U54 CA151880 · United States
NIA NIH HHS · R01AG17518 · United States
External Links
PubMed source DOI: 10.1128/MCB.01149-10 PMC Full Text View journal details
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