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Published 17 February 2004. doi:10.1083/jcb.200307081
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 164, Number 4, 593-602
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Article

Phosphorylation of paxillin by p38MAPK is involved in the neurite extension of PC-12 cells



Cai Huang1, Christoph H. Borchers2,3, Michael D. Schaller1,3,4, and Ken Jacobson1,3,4

1 Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, NC 27599
2 Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599
3 Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599
4 Comprehensive Center for Inflammatory Disorders, University of North Carolina, Chapel Hill, NC 27599

Address correspondence to Ken Jacobson, Dept. of Cell and Developmental Biology, University of North Carolina, 108 Taylor Hall, CB7090 Chapel Hill, NC 27599-7090. Tel.: (919) 966-5703. Fax: (919) 966-1856. email: frap{at}med.unc.edu

Cell adhesions play an important role in neurite extension. Paxillin, a focal adhesion adaptor protein involved in focal adhesion dynamics, has been demonstrated to be required for neurite outgrowth. However, the molecular mechanism by which paxillin regulates neurite outgrowth is unknown. Here, we show that paxillin is phosphorylated by p38MAPK in vitro and in nerve growth factor (NGF)–induced PC-12 cells. Ser 85 (Ser 83 for endogenous paxillin) is identified as one of major phosphorylation sites by phosphopeptide mapping and mass spectrometry. Moreover, expression of the Ser 85 -> Ala mutant of paxillin (paxS85A) significantly inhibits NGF-induced neurite extension of PC-12 cells, whereas expression of wild-type (wt) paxillin does not influence neurite outgrowth. Further experiments indicate that cells expressing paxS85A exhibit small, clustered focal adhesions which are not normally seen in cells expressing wt paxillin. Although wt paxillin and paxS85A have the same ability to bind vinculin and focal adhesion kinase, wt paxillin more efficiently associates with Pyk2 than paxS85A. Thus, phosphorylation of paxillin is involved in NGF-induced neurite extension of PC-12 cells, probably through regulating focal adhesion organization.

Key Words: neurons; nerve growth factor; focal adhesions; neurite outgrowth; mass spectrometry

The online version of this article contains supplemental material.

Abbreviations used in this paper: 2-D, 2-dimensional; JNK, c-jun NH2 terminus kinase; wt, wild-type.


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