A predominantly nuclear protein affecting cytoplasmic localization of {beta}-actin mRNA in fibroblasts and neurons

doi:10.1083/jcb.200105133

Published 7 January 2002. doi:10.1083/jcb.200105133

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© The Rockefeller University Press, 0021-9525/2002/1/41 $5.00
The Journal of Cell Biology, Volume 156, Number 1, January 7, 2002 41-52

Article

A predominantly nuclear protein affecting cytoplasmic localization of ß-actin mRNA in fibroblasts and neurons

Wei Gu¹, Feng Pan¹, Honglai Zhang², Gary J. Bassell² and Robert H. Singer¹

¹ Departments of Anatomy and Structural Biology and Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461
² Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461

Address correspondence to Robert H. Singer, Dept. of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Tel.: (718) 430-8646. Fax: (718) 430-8697. E-mail: rhsinger{at}aecom.yu.edu

The localization of ß-actin mRNA to the leading lamellaeof chicken fibroblasts and neurite growth cones of developingneurons requires a 54-nt localization signal (the zipcode) withinthe 3' untranslated region. In this study we have identifiedand isolated five proteins binding to the zipcode. One of thesewe previously identified as zipcode binding protein (ZBP)1,a 4-KH domain protein. A second is now investigated in detail:a 92-kD protein, ZBP2, that is especially abundant in extractsfrom embryonic brain. We show that ZBP2 is a homologue of thehuman hnRNP protein, KSRP, that appears to mediate pre-mRNAsplicing. However, ZBP2 has a 47–amino acid (aa) sequencenot present in KSRP. Various portions of ZBP2 fused to GFP indicatethat the protein most likely shuttles between the nucleus andthe cytoplasm, and that the 47-aa insert promotes the nuclearlocalization. Expression of a truncated ZBP2 inhibits the localizationof ß-actin mRNA in both fibroblast and neurons. Thesedata suggest that ZBP2, although predominantly a nuclear protein,has a role in the cytoplasmic localization of ß-actinmRNA.

Key Words: RNA localization; RNA binding proteins; nuclear-cytoplasmic trafficking; RNA splicing; KH domain proteins

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