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PMID: 15302913 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

RNA recognition motif 2 directs the recruitment of SF2/ASF to nuclear stress bodies.

Nucleic acids research ·Vol. 32 ·No. 14 ·2004-00-00 ·Pages 4127-36

Chiodi I, Corioni M, Giordano M, Valgardsdottir R, Ghigna C, Cobianchi F, Xu RM, Riva S, Biamonti G

Abstract

Heat shock induces the transcriptional activation of large heterochromatic regions of the human genome composed of arrays of satellite III DNA repeats. A number of RNA-processing factors, among them splicing factor SF2/ASF, associate with these transcription factors giving rise to nuclear stress bodies (nSBs). Here, we show that the recruitment of SF2/ASF to these structures is mediated by its second RNA recognition motif. Amino acid substitutions in the first alpha-helix of this domain, but not in the beta-strand regions, abrogate the association with nSBs. The same mutations drastically affect the in vivo activity of SF2/ASF in the alternative splicing of adenoviral E1A transcripts. Sequence analysis identifies four putative high-affinity binding sites for SF2/ASF in the transcribed strand of the satellite III DNA. We have verified by gel mobility shift assays that the second RNA-binding domain of SF2/ASF binds at least one of these sites. Our analysis suggests that the recruitment of SF2/ASF to nSBs is mediated by a direct interaction with satellite III transcripts and points to the second RNA-binding domain of the protein as the major determinant of this interaction.

MeSH Terms
Alternative Splicing Amino Acid Motifs Amino Acid Sequence Amino Acid Substitution Binding Sites Cell Nucleus Structures/metabolism DNA, Satellite/chemistry,metabolism HeLa Cells Heat-Shock Response Humans Molecular Sequence Data Nuclear Proteins/chemistry,metabolism Protein Transport RNA-Binding Proteins/chemistry,metabolism Serine-Arginine Splicing Factors Two-Hybrid System Techniques
Chemicals
DNA, Satellite Nuclear Proteins RNA-Binding Proteins Serine-Arginine Splicing Factors
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Chiodi Ilaria
Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche, Via Abbiategrasso 207, 27100 Pavia, Italy.
Corioni Margherita
Giordano Manuela
Valgardsdottir Rut
Ghigna Claudia
Cobianchi Fabio
Xu Rui-Ming
Riva Silvano
Biamonti Giuseppe
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkh759
Published
2004-00-00
Epub
2004-00-09
Pages
4127-36
Language
English
Region
England
NLM ID
0411011
PMCID
PMC514380
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkh759 PMC Full Text View journal details
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