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PMID: 11739736 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Characterization of U2AF(6), a splicing factor related to U2AF(35).

Molecular and cellular biology ·Vol. 22 ·No. 1 ·2002-01-00 ·Pages 221-30

Shepard J, Reick M, Olson S, Graveley BR

Abstract

The essential splicing factor U2AF (U2 auxiliary factor) is a heterodimer composed of 65-kDa (U2AF(65)) and 35-kDa (U2AF(35)) subunits. U2AF(35) has multiple functions in pre-mRNA splicing. First, U2AF(35) has been shown to function by directly interacting with the AG at the 3' splice site. Second, U2AF(35) is thought to play a role in the recruitment of U2AF(65) by serine-arginine-rich (SR) proteins in enhancer-dependent splicing. It has been proposed that the physical interaction between the arginine-serine-rich (RS) domain of U2AF(35) and SR proteins is important for this activity. However, other data suggest that this may not be the case. Here, we report the identification of a mammalian gene that encodes a 26-kDa protein bearing strong sequence similarity to U2AF(35), designated U2AF(26). The N-terminal 187 amino acids of U2AF(35) and U2AF(26) are nearly identical. However, the C-terminal domain of U2AF(26) lacks many characteristics of the U2AF(35) RS domain and, therefore, might be incapable of interacting with SR proteins. We show that U2AF(26) can associate with U2AF(65) and can functionally substitute for U2AF(35) in both constitutive and enhancer-dependent splicing, demonstrating that the RS domain of the small U2AF subunit is not required for splicing enhancer function. Finally, we show that U2AF(26) functions by enhancing the binding of U2AF(65) to weak 3' splice sites. These studies identify U2AF(26) as a mammalian splicing factor and demonstrate that distinct U2AF complexes can participate in pre-mRNA splicing. Based on its sequence and functional similarity to U2AF(35), U2AF(26) may play a role in regulating alternative splicing.

MeSH Terms
Amino Acid Sequence Animals Cell Fractionation Cloning, Molecular HeLa Cells Humans Immunohistochemistry Mice Molecular Sequence Data Nuclear Proteins Protein Binding Protein Structure, Tertiary Protein Subunits RNA Splice Sites/genetics RNA Splicing/physiology Ribonucleoproteins/chemistry,genetics,isolation & purification,metabolism Sequence Alignment Splicing Factor U2AF Tissue Distribution
Chemicals
Nuclear Proteins Protein Subunits RNA Splice Sites Ribonucleoproteins Splicing Factor U2AF U2AF2 protein, human U2af1l4 protein, mouse Zrsr1 protein, mouse
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Shepard Jeremiah
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
Reick Martin
Olson Sara
Graveley Brenton R
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.22.1.221-230.2002
Published
2002-01-00
Pages
221-30
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC134218
Subset
IM
Grants
NIGMS NIH HHS · R01 GM062516 · United States
NIMH NIH HHS · R37 MH059388 · United States
NIMH NIH HHS · 5 R37 MH59388-03 · United States
NIGMS NIH HHS · R01 GM62516-01 · United States
External Links
PubMed source DOI: 10.1128/MCB.22.1.221-230.2002 PMC Full Text View journal details
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