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A correction to this article has been published: J. Cell Biol. 137 (7) 1683
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© The Rockefeller University Press, 0021-9525/1997//1263 $5.00
The Journal of Cell Biology, Volume 136, Number 6, , 1997 1263-1270

Article

β-Actin Messenger RNA Localization and Protein Synthesis Augment Cell Motility



Edward H. Kislauskis*, Xiao-chun Zhu*, and Robert H. Singer{ddagger}

* Department of Cell Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655; and {ddagger} Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461

In chicken embryo fibroblasts (CEFs), β-actin mRNA localizes near an actin-rich region of cytoplasm specialized for motility, the lamellipodia. This localization is mediated by isoform-specific 3'-untranslated sequences (zipcodes) and can be inhibited by antizipcode oligodeoxynucleotides (ODNs) (Kislauskis, E.H., X.-C. Zhu, and R.H. Singer. 1994. J. Cell Biol. 127: 441–451). This inhibition of β-actin mRNA localization resulted in the disruption of fibroblast polarity and, presumably, cell motility. To investigate the role of β-actin mRNA in motility, we correlated time-lapse images of moving CEFs with the distribution of β-actin mRNA in these cells. CEFs with localized β-actin mRNA moved significantly further over the same time period than did CEFs with nonlocalized mRNA. Antizipcode ODN treatment reduced this cell translocation while control ODN treatments showed no effect. The temporal relationship of β-actin mRNA localization to cell translocation was investigated using serum addition to serum-deprived cultures. β-actin mRNA was not localized in serum-deprived cells but became localized within minutes after serum addition (Latham, V.M., E.H. Kislauskis, R.H. Singer, and A.F. Ross. 1994. J. Cell Biol. 126:1211–1219). Cell translocation increased over the next 90 min, and actin synthesis likewise increased. Puromycin reduced this cell translocation and blocked this induction in cytosolic actin content. The serum induction of cell movement was also inhibited by antizipcode ODNs. These observations support the hypothesis that β-actin mRNA localization and consequent protein synthesis augment cell motility.

Abbreviations used in this paper: CEF, chicken embryo fibroblasts; ODN, oligodeoxynucleotides.

Essential conceptual contributions were made by John Condeelis (Albert Einstein College of Medicine), and critical comments on early drafts were made by Yu-Li Wang (Worcester Foundation for Biomedical Research). The authors thank Dr. Peter Quesenberry for access to his videomicroscope. Helpful comments related to this manuscript also were made by members of the laboratory: Tony Ross, Joan Politz, Krishan Taneja, and Chris Powers. We appreciate Birgit Koppetsch's skill at preparing primary CEF cultures and the secretarial help provided by Terri O'Toole. The essence of much of this work was published in abstract form (1995. Mol. Biol. Cell. 6[Suppl.]:309a). Funding for this work was provided to E.H. Kislauskis by a Muscular Dystrophy Research Grant and to R.H. Singer from National Institutes of Health (grant AR41480).

Address all correspondence to Edward H. Kislauskis, Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Avenue, Worcester, MA 01655. Tel.: (508) 856-4230. Fax: (508) 856-5612. E-mail: ehk{at}insitu.ummed.edu


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