Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Subcellular targeting of oxidants during endothelial cell migration

Subcellular targeting of oxidants during endothelial cell migration Endogenous oxidants participate in endothelial cell migration, suggesting that the enzymatic source of oxidants, like other proteins controlling cell migration, requires precise subcellular localization for spatial confinement of signaling effects. We found that the nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase adaptor p47 phox and its binding partner TRAF4 were sequestered within nascent, focal complexlike structures in the lamellae of motile endothelial cells. TRAF4 directly associated with the focal contact scaffold Hic-5, and the knockdown of either protein, disruption of the complex, or oxidant scavenging blocked cell migration. An active mutant of TRAF4 activated the NADPH oxidase downstream of the Rho GTPases and p21-activated kinase 1 (PAK1) and oxidatively modified the focal contact phosphatase PTP-PEST. The oxidase also functioned upstream of Rac1 activation, suggesting its participation in a positive feedback loop. Active TRAF4 initiated robust membrane ruffling through Rac1, PAK1, and the oxidase, whereas the knockdown of PTP-PEST increased ruffling independent of oxidase activation. Our data suggest that TRAF4 specifies a molecular address within focal complexes that is targeted for oxidative modification during cell migration. Footnotes Abbreviations used in this paper: 5′-IAF, 5′-iodoacetamidofluorescein; CRIB, Cdc42–Rac1 interaction binding; HUVEC, human umbilical vein endothelial cell; JNK, c-Jun NH 2 -terminal kinase; MOI, multiplicity of infection; PAK, p21-activated kinase; PID, PAK inhibitory domain; siRNA, short inhibitory RNA; TIRF, total internal reflection fluorescence. Submitted: 1 July 2005 Accepted: 3 November 2005 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Journal of Cell Biology Rockefeller University Press

Subcellular targeting of oxidants during endothelial cell migration

Loading next page...
 
/lp/rockefeller-university-press/subcellular-targeting-of-oxidants-during-endothelial-cell-migration-2krVJxkOES

References

References for this paper are not available at this time. We will be adding them shortly, thank you for your patience.

Publisher
Rockefeller University Press
Copyright
Copyright © 2005, by The Rockefeller University Press
ISSN
0021-9525
eISSN
1540-8140
DOI
10.1083/jcb.200507004
pmid
16330715
Publisher site
See Article on Publisher Site

Abstract

Endogenous oxidants participate in endothelial cell migration, suggesting that the enzymatic source of oxidants, like other proteins controlling cell migration, requires precise subcellular localization for spatial confinement of signaling effects. We found that the nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase adaptor p47 phox and its binding partner TRAF4 were sequestered within nascent, focal complexlike structures in the lamellae of motile endothelial cells. TRAF4 directly associated with the focal contact scaffold Hic-5, and the knockdown of either protein, disruption of the complex, or oxidant scavenging blocked cell migration. An active mutant of TRAF4 activated the NADPH oxidase downstream of the Rho GTPases and p21-activated kinase 1 (PAK1) and oxidatively modified the focal contact phosphatase PTP-PEST. The oxidase also functioned upstream of Rac1 activation, suggesting its participation in a positive feedback loop. Active TRAF4 initiated robust membrane ruffling through Rac1, PAK1, and the oxidase, whereas the knockdown of PTP-PEST increased ruffling independent of oxidase activation. Our data suggest that TRAF4 specifies a molecular address within focal complexes that is targeted for oxidative modification during cell migration. Footnotes Abbreviations used in this paper: 5′-IAF, 5′-iodoacetamidofluorescein; CRIB, Cdc42–Rac1 interaction binding; HUVEC, human umbilical vein endothelial cell; JNK, c-Jun NH 2 -terminal kinase; MOI, multiplicity of infection; PAK, p21-activated kinase; PID, PAK inhibitory domain; siRNA, short inhibitory RNA; TIRF, total internal reflection fluorescence. Submitted: 1 July 2005 Accepted: 3 November 2005

Journal

The Journal of Cell BiologyRockefeller University Press

Published: Dec 5, 2005

There are no references for this article.