Nucleic Acids Research, 1994, Vol. 22, No. 2 238-246
© 1994
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AU-A, an RNA-binding activity distinct from hnRNP A1, is selective for AUUUA repeats and shuttles between the nucleus and the cytoplasm
Departments of Molecular Genetics and Cell Biology, and Medicine, and Howard Hughes Medical Institute, University of Chicago 5841 S. Maryland, MC 1028, Chicago, IL 60637 1Department of Biological Chemistry, Johns Hopkins University, School of Medicine Baltimore, MD 21205, USA
*To whom correspondence should be addressed
Received August 9, 1993. Accepted December 6, 1993.
The 3'-untranslated regions of many labile transcripts contain AU-rich sequences that serve as cis determinants of mRNA stability and translatlonal efficiency. Using a photocrossllnklng technique, our laboratory has previously defined three cytoplasmlc RNA-bindlng activities specific for the AUUUA multimers found in the 3'-untranslated regions of lymphokine mRNAs. One of these activities, AU-A, has an apparent molecular mass of 34 kDa, Is constltutively expressed In both primary T cells and the Jurkat T cell leukemia line, and binds to a variety of U-rich RNA sequences. Previous studies had shown that AU-A is more prevalent In the nucleus than the cytoplasm, raising the possibility that AU-A is really a nuclear RNA-blnding activity that is found In cytoplasmlc extracts because of nuclear leakage during cell fractionatlon. We now show that AU-A shuttles between the cytoplasm and the nucleus. Our results indicate that AU-A Is a candidate protein component of ribonucleoprotein complexes that participate in nucleocytoplasmic transport of mRNA and cytoplasmic mRNA metabolism. The properties of AU-A activity are similar to those of heterogenous nuclear ribonucleoprotein A1 (hnRNP A1). However, using monoclonal antibodies to hnRNP A1 and protease digestion patterns, we show that AU-A activity and hnRNP A1 protein are distinct. These studies have also allowed us to define a fourth RNA-binding activity of apparent molecular mass 41 kDa with specificity for AUUUA multimers. This activity is restricted to the nucleus and contains the hnRNP C protein.
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