The focal adhesion-associated proteins DOCK5 and GIT2 comprise a rheostat in control of epithelial invasion.
DOCK proteins are guanine nucleotide exchange factors for Rac and Cdc42 GTPases. DOCK1 is the founding member of the family and acts downstream of integrins via the canonical Crk-p130Cas complex to activate Rac GTPases in numerous contexts. In contrast, DOCK5, which possesses the greatest similarity to DOCK1, remains sparingly studied. Here we establish that DOCK5 has a non-redundant role in regulating motile and invasive capacities of epithelial cells. DOCK1 is constitutively associated with sites of integrin attachment termed focal adhesions (FAs). In contrast, we demonstrate that DOCK5 recruitment to FAs in Hela cells is restricted by GIT2, an established regulator of FA signaling. We determine that GIT2 is targeted to FAs in response to Rho-ROCK signaling and actomyosin contractility. Accordingly, inhibition of ROCK activity or MLC function promotes enrichment of DOCK5 in membrane protrusions and nascent cell-substratum adhesions. We further demonstrate that GIT2 inhibits the interaction of DOCK5 with Crk. Moreover, we show that depletion of GIT2 promotes DOCK5-dependent activation of the Crk-p130Cas signaling cascade to promote Rac1-mediated lamellipodial protrusion and FA turnover. The antagonism between GIT2 and DOCK5 extends to non-transformed MCF10A mammary epithelial cells, with DOCK5 'dialing-up' and GIT2 'dialing-down' invasiveness. Finally, we determine that DOCK5 inhibition attenuates invasion and metastasis of MDA-MB-231 cells and prolongs life span of mice injected with these cells. Collectively, our work identifies DOCK5 as a key regulator of epithelial invasion and metastasis, and demonstrates that suppression of DOCK5 by GIT2 represents a previously unappreciated mechanism for coordination of Rho and Rac GTPases.
GI Cell Biology Research Laboratory, Children's Hospital Boston and Harvard Medical School, Boston, MA, USA.
S R
SR
Frank
GI Cell Biology Research Laboratory, Children's Hospital Boston and Harvard Medical School, Boston, MA, USA.
C P
CP
Köllmann
GI Cell Biology Research Laboratory, Children's Hospital Boston and Harvard Medical School, Boston, MA, USA.
J F
JF
van Lidth de Jeude
GI Cell Biology Research Laboratory, Children's Hospital Boston and Harvard Medical School, Boston, MA, USA.
J R
JR
Thiagarajah
Finsen Laboratory, Rigshospitalet, Copenhagen Biocenter, Copenhagen, Denmark.
L H
LH
Engelholm
Biotech Research and Innovation Centre, University of Copenhagen, Copenhagen, Denmark.
M
M
Frödin
GI Cell Biology Research Laboratory, Children's Hospital Boston and Harvard Medical School, Boston, MA, USA.
S H
SH
Hansen
D000209:Acute-Phase Proteins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D020690:GTPase-Activating Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics* / Q000378:metabolism; D006367:HeLa Cells; D064593:Heterografts; D006801:Humans; D051379:Mice; D008954:Models, Biological; D009362:Neoplasm Metastasis; D011485:Protein Binding; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000235:genetics
D000209:Acute-Phase Proteins; C578026:Dock5 protein, human; C430213:GIT2 protein, human; D020690:GTPase-Activating Proteins; D020662:Guanine Nucleotide Exchange Factors; D034741:RNA, Small Interfering; C010781:acute-phase protein rho
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