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A more recent version of this article appeared on February 1, 2003
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Submitted on June 17, 2002
Revised on October 2, 2002
Accepted on October 21, 2002
1 School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom
2 MRC Laboratory of Molecular and Cellular Biology, University College London, London WC1E 6BT, United Kingdom
3 School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom (present address: Faculty of Medicine and Biological Sciences, University of Leicester, Leicester LE1 9HN, United Kingdom)
* Corresponding author. E-mail address: l.m.machesky{at}bham.ac.uk.
Cell motility and cell polarity are essential for morphogenesis, immune system function and tissue repair. Many animal cells move by crawling and one main driving force for movement is derived from the co-ordinated assembly and disassembly of actin filaments. As tissue culture cells migrate to close a scratch wound, this directional extension is accompanied by Golgi apparatus reorientation, to face the leading wound edge, giving the motile cell inherent polarity aligned relative to the wound edge and to the direction of cell migration. Cellular proteins essential for actin polymerization downstream of Rho family GTPases include the Arp2/3 complex as an actin nucleator and members of the Wiskott-Aldrich Syndrome protein (WASP) family as activators of the Arp2/3 complex. We therefore analyzed the involvement of the Arp2/3 complex and WASP-family proteins in in vitro wound healing assays using NIH 3T3 fibroblasts and astrocytes. In NIH 3T3 cells, we found that actin and Arp2/3 complex contributed to cell polarity establishment. Moreover, overexpression of N-terminal fragments of Scar2 (but not N-WASP or Scar1 or Scar3) interfere with NIH 3T3 Golgi polarization, but not with cell migration. In contrast, actin, Arp2/3 and WASP-family proteins did not appear to be involved in Golgi polarization in astrocytes. Our results thus indicate that the requirement for Golgi polarity establishment is cell-type specific. Furthermore, in NIH 3T3 cells, Scar2 and the Arp2/3 complex appear to be involved in the establishment and maintenance of Golgi polarity during directed migration.
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