TY - JOUR
T1 - Molecular mechanisms regulating the subcellular localization of p95-APP1 between the endosomal recycling compartment and sites of actin organization at the cell surface
AU - Matafora, Vittoria
AU - Paris, Simona
AU - Dariozzi, Simona
AU - de Curtis, Ivan
PY - 2001
Y1 - 2001
N2 - Cell migration requires coordination between adhesion, actin organization and membrane traffic. Rac and ARF6 have been shown to cooperate for the organization of actin at the cell surface. Recently, the GIT family of ARF-GAPs has been identified, which includes proteins that can functionally interact with both ARF and Rac GTPases. The p95-APP1 protein is a member of this family, isolated as part of a multi-molecular complex interacting with GTP-Rac. Our previous work has indicated that this protein may be part of the machinery redirecting membrane recycling towards sites of protrusion during locomotion. By analyzing the distribution and the effects of truncated forms of p95-APP1, we show here that the lack of the ARF-GAP domain of p95-APP1 dramatically shifts its localization to large vesicles. The use of several markers of the endocytic pathway has revealed that the truncated p95-APP1 localizes specifically to a Rab11-, transferrin receptor-positive compartment. Other markers are excluded from the p95-APP1-positive vesicles, while known components of the multi-molecular complex colocalize with truncated p95-APP1 in this compartment. Coexpression of a constitutively active form of Rac induces the redistribution of the truncated constructs and of the associated PIX, PAK, and paxillin to peripheral sites of Rac-mediated actin organization, and the disassembly of the large Rab11-positive vesicles. Together, the data presented indicate that p95-APP1 is part of a complex that shuttles between the plasma membrane and the endocytic recycling compartment, and suggest that the dynamic redistribution of the p95-APP1-containing complex is mediated both by the ARF-GAP domain, and by the recruitment of the complex at the cell surface at sites of Rac activation.
AB - Cell migration requires coordination between adhesion, actin organization and membrane traffic. Rac and ARF6 have been shown to cooperate for the organization of actin at the cell surface. Recently, the GIT family of ARF-GAPs has been identified, which includes proteins that can functionally interact with both ARF and Rac GTPases. The p95-APP1 protein is a member of this family, isolated as part of a multi-molecular complex interacting with GTP-Rac. Our previous work has indicated that this protein may be part of the machinery redirecting membrane recycling towards sites of protrusion during locomotion. By analyzing the distribution and the effects of truncated forms of p95-APP1, we show here that the lack of the ARF-GAP domain of p95-APP1 dramatically shifts its localization to large vesicles. The use of several markers of the endocytic pathway has revealed that the truncated p95-APP1 localizes specifically to a Rab11-, transferrin receptor-positive compartment. Other markers are excluded from the p95-APP1-positive vesicles, while known components of the multi-molecular complex colocalize with truncated p95-APP1 in this compartment. Coexpression of a constitutively active form of Rac induces the redistribution of the truncated constructs and of the associated PIX, PAK, and paxillin to peripheral sites of Rac-mediated actin organization, and the disassembly of the large Rab11-positive vesicles. Together, the data presented indicate that p95-APP1 is part of a complex that shuttles between the plasma membrane and the endocytic recycling compartment, and suggest that the dynamic redistribution of the p95-APP1-containing complex is mediated both by the ARF-GAP domain, and by the recruitment of the complex at the cell surface at sites of Rac activation.
KW - Actin organization
KW - Cell motility
KW - GTP-binding proteins
KW - Recycling endosomes
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UR - http://www.scopus.com/inward/citedby.url?scp=0035694169&partnerID=8YFLogxK
M3 - Article
C2 - 11792816
AN - SCOPUS:0035694169
VL - 114
SP - 4509
EP - 4520
JO - Journal of Cell Science
JF - Journal of Cell Science
SN - 0021-9533
IS - 24
ER -