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J. Biol. Chem., Vol. 276, Issue 47, 43850-43859, November 23, 2001
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From the Department of Molecular, Cellular, and Developmental
Biology and Center for Molecular Biology of RNA, Sinsheimer
Laboratories, University of California,
Santa Cruz, California 95064
Members of the heterogeneous nuclear
ribonucleoprotein (hnRNP) H protein family, H, H', F, and 2H9, are
involved in pre-mRNA processing. We analyzed the assembly of these
proteins from splicing extracts onto four RNA regulatory elements as
follows: a high affinity hnRNP A1-binding site (WA1), a sequence
involved in Rev-dependent export (p17gag INS), an
exonic splicing silencer from the
Determination of the RNA Binding Specificity of the Heterogeneous
Nuclear Ribonucleoprotein (hnRNP) H/H'/F/2H9 Family*
-tropomyosin gene, and an intronic
splicing regulator (downstream control sequence (DCS) from the
c-src gene. The entire family binds the WA1, instability (INS), and -tropomyosin substrates, and the core-binding site for
each is a run of three G residues followed by an A. Transfer of small
regions containing this sequence to a substrate lacking hnRNP H binding
activity is sufficient to promote binding of all family members. The
c-src DCS has been shown to assemble hnRNP H, not hnRNP F,
from HeLa cell extracts, and we show that hnRNP 2H9 does not bind this
element. The DCS contains five G residues followed by a C. Mutation of
the C to an A changes the specificity of the DCS from a substrate that
binds only hnRNP H/H' to a binding site for all hnRNP H family members.
We conclude that the sequence GGGA is recognized by all hnRNP H family proteins.
*
This work was supported by Grant R99-SC-085 from the
University of California University-wide AIDS Research Program and
NIGMS Grant 1R01GM61646 from the National Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 831-459-5131;
Fax: 831-459-3737; E-mail: zahler@biology.ucsc.edu.
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