TY - JOUR
T1 - Imaging sites of N-WASP activity in lamellipodia and invadopodia of carcinoma cells
AU - Lorenz, Mike
AU - Yamaguchi, Hideki
AU - Wang, Yarong
AU - Singer, Robert H.
AU - Condeelis, John
N1 - Funding Information:
We thank M. Way (Cancer Research UK, London, UK) for his generous gift of the rat GFP-N-WASP plasmid and M. Cammer (Analytical Imaging Facility, AECOM) for his help with the confocal microscopy. This work was supported by grants from the National Institutes of Health to J.C. (38511) and R.H.S.
PY - 2004/4/20
Y1 - 2004/4/20
N2 - Cell migration is crucial for many biological and pathological processes such as chemotaxis of immune cells, fibroblast migration during wound healing, and tumor cell invasion and metastasis. Cells migrate forward by extending membrane protrusions. The formation of these protrusions is driven by assembly of actin filaments at the leading edge [1]. Neural Wiskott-Aldrich syndrome protein (N-WASP), a ubiquitous member of the WASP family, induces actin polymerization by activating Arp2/3 complex and is thought to regulate the formation of membrane protrusions [2, 3]. However, it is totally unclear how N-WASP activity is spatially and temporally regulated inside migrating cells. To detect and image sites of N-WASP activity during cell motility and invasion in carcinoma cells, we designed an N-WASP fluorescence resonance energy transfer (FRET) biosensor that distinguishes between the active and inactive conformations and mimics the function of endogenous N-WASP. Our data show that N-WASP is involved in lamellipodia extension, where it is activated at the leading edge, as well as in invadopodia formation of invasive carcinoma cells, where it is activated at the base. This is the first time that the activity of full-length N-WASP has been visualized in vivo, and this has lead to new insights for N-WASP function.
AB - Cell migration is crucial for many biological and pathological processes such as chemotaxis of immune cells, fibroblast migration during wound healing, and tumor cell invasion and metastasis. Cells migrate forward by extending membrane protrusions. The formation of these protrusions is driven by assembly of actin filaments at the leading edge [1]. Neural Wiskott-Aldrich syndrome protein (N-WASP), a ubiquitous member of the WASP family, induces actin polymerization by activating Arp2/3 complex and is thought to regulate the formation of membrane protrusions [2, 3]. However, it is totally unclear how N-WASP activity is spatially and temporally regulated inside migrating cells. To detect and image sites of N-WASP activity during cell motility and invasion in carcinoma cells, we designed an N-WASP fluorescence resonance energy transfer (FRET) biosensor that distinguishes between the active and inactive conformations and mimics the function of endogenous N-WASP. Our data show that N-WASP is involved in lamellipodia extension, where it is activated at the leading edge, as well as in invadopodia formation of invasive carcinoma cells, where it is activated at the base. This is the first time that the activity of full-length N-WASP has been visualized in vivo, and this has lead to new insights for N-WASP function.
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U2 - 10.1016/j.cub.2004.04.008
DO - 10.1016/j.cub.2004.04.008
M3 - Article
C2 - 15084285
AN - SCOPUS:1942508987
SN - 0960-9822
VL - 14
SP - 697
EP - 703
JO - Current Biology
JF - Current Biology
IS - 8
ER -