XB-ART-37817
Dev Biol
July 1, 2008;
319
(1):
86-99.
PACSIN2 regulates cell adhesion during gastrulation in Xenopus laevis.
Abstract
We previously identified the adaptor protein PACSIN2 as a negative regulator of ADAM13 proteolytic function. In Xenopus embryos, PACSIN2 is ubiquitously expressed, suggesting that PACSIN2 may control other proteins during development. To investigate this possibility, we studied PACSIN2 function during Xenopus gastrulation and in XTC cells. Our results show that PACSIN2 is localized to the plasma membrane via its coiled-coil domain. We also show that increased levels of PACSIN2 in embryos inhibit gastrulation, fibronectin (FN) fibrillogenesis and the ability of ectodermal cells to spread on a FN substrate. These effects require PACSIN2 coiled-coil domain and are not due to a reduction of FN or integrin expression and/or trafficking. The expression of a Mitochondria Anchored PACSIN2 (PACSIN2-MA) sequesters wild type PACSIN2 to mitochondria, and blocks gastrulation without interfering with cell spreading or FN fibrillogenesis but perturbs both epiboly and convergence/extension. In XTC cells, the over-expression of PACSIN2 but not PACSIN2-MA prevents the localization of integrin beta1 to focal adhesions (FA) and filamin to stress fiber. PACSIN2-MA prevents filamin localization to membrane ruffles but not to stress fiber. We propose that PACSIN2 may regulate gastrulation by controlling the population of activated alpha5beta1 integrin and cytoskeleton strength during cell movement.
PubMed ID: 18495106
PMC ID: PMC2494715
Article link: Dev Biol
Grant support: [+]
DE016289 NIDCR NIH HHS, DE14365 NIDCR NIH HHS, R-01HD26402 NICHD NIH HHS , R01 DE014365-01 NIDCR NIH HHS, R01 DE014365-02 NIDCR NIH HHS, R01 DE014365-03 NIDCR NIH HHS, R01 DE014365-04 NIDCR NIH HHS, R01 DE014365-05 NIDCR NIH HHS, R01 DE016289-01A2 NIDCR NIH HHS, R01 DE016289-02 NIDCR NIH HHS, R01 DE016289-03 NIDCR NIH HHS, R01 HD026402-10 NICHD NIH HHS , R01 HD026402-11 NICHD NIH HHS , R01 HD026402-12 NICHD NIH HHS , R01 HD026402-13 NICHD NIH HHS , R01 DE016289 NIDCR NIH HHS, R01 DE014365 NIDCR NIH HHS, R01 HD026402 NICHD NIH HHS
Species referenced: Xenopus
Genes referenced: acta4 actl6a adam33 bcr chrd.1 flna fn1 gal.2 itga5 itgav itgb1 myc pacsin2 tbx2 tbxt tubb2b
Antibodies: Adam33 Ab1 Flna Ab1 Fluro-phalloidin Ab Fn1 Ab1 Fn1 Ab3 Itgav Ab1 Itgb1 Ab1 Itgb1 Ab3 Mitochondria Lectin Myc Ab5 Pacsin2 Ab1
Morpholinos: pacsin2 MO1 pacsin2 MO2
Article Images: [+] show captions
References [+] :
Alfandari,
Xenopus ADAM 13 is a metalloprotease required for cranial neural crest-cell migration.
2001, Pubmed,
Xenbase
Alfandari,
Xenopus ADAM 13 is a metalloprotease required for cranial neural crest-cell migration.
2001,
Pubmed
,
Xenbase
Alfandari, Integrin alpha5beta1 supports the migration of Xenopus cranial neural crest on fibronectin. 2003, Pubmed , Xenbase
Alfandari, Integrin alpha v subunit is expressed on mesodermal cell surfaces during amphibian gastrulation. 1995, Pubmed , Xenbase
Alfandari, ADAM 13: a novel ADAM expressed in somitic mesoderm and neural crest cells during Xenopus laevis development. 1997, Pubmed , Xenbase
Badour, The Wiskott-Aldrich syndrome protein acts downstream of CD2 and the CD2AP and PSTPIP1 adaptors to promote formation of the immunological synapse. 2003, Pubmed
Boucaut, Prevention of gastrulation but not neurulation by antibodies to fibronectin in amphibian embryos. 1984, Pubmed
Boucaut, Biologically active synthetic peptides as probes of embryonic development: a competitive peptide inhibitor of fibronectin function inhibits gastrulation in amphibian embryos and neural crest cell migration in avian embryos. 1984, Pubmed
Bubeck, Ligand recruitment by vinculin domains in transfected cells. 1997, Pubmed
Calderwood, Talin controls integrin activation. 2004, Pubmed
Chitu, Pombe Cdc15 homology (PCH) proteins: coordinators of membrane-cytoskeletal interactions. 2007, Pubmed
Cohen, Active zones on motor nerve terminals contain alpha 3beta 1 integrin. 2000, Pubmed , Xenbase
Cousin, PACSIN2 is a regulator of the metalloprotease/disintegrin ADAM13. 2000, Pubmed , Xenbase
Cousin, A PTP-PEST-like protein affects alpha5beta1-integrin-dependent matrix assembly, cell adhesion, and migration in Xenopus gastrula. 2004, Pubmed , Xenbase
Cunningham, Actin-binding protein requirement for cortical stability and efficient locomotion. 1992, Pubmed
Darribère, Distinct regions of human fibronectin are essential for fibril assembly in an in vivo developing system. 1992, Pubmed
Darribère, In vivo analyses of integrin beta 1 subunit function in fibronectin matrix assembly. 1990, Pubmed
Davidson, Mesendoderm extension and mantle closure in Xenopus laevis gastrulation: combined roles for integrin alpha(5)beta(1), fibronectin, and tissue geometry. 2002, Pubmed , Xenbase
Davidson, Integrin alpha5beta1 and fibronectin regulate polarized cell protrusions required for Xenopus convergence and extension. 2006, Pubmed , Xenbase
Fankhauser, The S. pombe cdc15 gene is a key element in the reorganization of F-actin at mitosis. 1995, Pubmed
Flanagan, Filamin A, the Arp2/3 complex, and the morphology and function of cortical actin filaments in human melanoma cells. 2001, Pubmed
Gaultier, ADAM13 disintegrin and cysteine-rich domains bind to the second heparin-binding domain of fibronectin. 2002, Pubmed , Xenbase
Gawantka, A beta 1-integrin associated alpha-chain is differentially expressed during Xenopus embryogenesis. 1995, Pubmed , Xenbase
Ghadimi, Identification of interaction partners of the cytosolic polyproline region of CD95 ligand (CD178). 2002, Pubmed
Ginsberg, Integrin regulation. 2005, Pubmed
Glogauer, The role of actin-binding protein 280 in integrin-dependent mechanoprotection. 1998, Pubmed
Halbach, PACSIN 1 forms tetramers via its N-terminal F-BAR domain. 2007, Pubmed
Hynes, Integrins: bidirectional, allosteric signaling machines. 2002, Pubmed
Jha, Evidence for the involvement of proline-directed serine/threonine phosphorylation in sperm capacitation. 2006, Pubmed
Joos, Integrin alpha 5 during early development of Xenopus laevis. 1995, Pubmed , Xenbase
Keller, Mechanisms of convergence and extension by cell intercalation. 2000, Pubmed
Kessels, The syndapin protein family: linking membrane trafficking with the cytoskeleton. 2004, Pubmed
Kessels, Syndapin oligomers interconnect the machineries for endocytic vesicle formation and actin polymerization. 2006, Pubmed
Kiema, The molecular basis of filamin binding to integrins and competition with talin. 2006, Pubmed
Lee, Temporal and spatial regulation of fibronectin in early Xenopus development. 1984, Pubmed , Xenbase
Li, A cdc15-like adaptor protein (CD2BP1) interacts with the CD2 cytoplasmic domain and regulates CD2-triggered adhesion. 1999, Pubmed
Marsden, Regulation of cell polarity, radial intercalation and epiboly in Xenopus: novel roles for integrin and fibronectin. 2001, Pubmed , Xenbase
Marsden, Integrin-ECM interactions regulate cadherin-dependent cell adhesion and are required for convergent extension in Xenopus. 2003, Pubmed , Xenbase
Modregger, All three PACSIN isoforms bind to endocytic proteins and inhibit endocytosis. 2001, Pubmed
Mori, PACSIN3 binds ADAM12/meltrin alpha and up-regulates ectodomain shedding of heparin-binding epidermal growth factor-like growth factor. 2003, Pubmed
Mould, Regulation of integrin alpha 5 beta 1-fibronectin interactions by divalent cations. Evidence for distinct classes of binding sites for Mn2+, Mg2+, and Ca2+. 1995, Pubmed
Na, Differential regulation of cell adhesive functions by integrin alpha subunit cytoplasmic tails in vivo. 2003, Pubmed , Xenbase
Nikki, FAP52 regulates actin organization via binding to filamin. 2002, Pubmed
Nikki, Focal adhesion protein FAP52 self-associates through a sequence conserved among the members of the PCH family proteins. 2002, Pubmed
Park, Subcellular localization and signaling properties of dishevelled in developing vertebrate embryos. 2005, Pubmed , Xenbase
Pistor, The ActA protein of Listeria monocytogenes acts as a nucleator inducing reorganization of the actin cytoskeleton. 1994, Pubmed
Qualmann, Syndapin isoforms participate in receptor-mediated endocytosis and actin organization. 2000, Pubmed
Ramos, Xenopus embryonic cell adhesion to fibronectin: position-specific activation of RGD/synergy site-dependent migratory behavior at gastrulation. 1996, Pubmed , Xenbase
Ramos, Integrin-dependent adhesive activity is spatially controlled by inductive signals at gastrulation. 1996, Pubmed , Xenbase
Ridley, Cell migration: integrating signals from front to back. 2003, Pubmed
Simpson, SH3-domain-containing proteins function at distinct steps in clathrin-coated vesicle formation. 1999, Pubmed
Smith, Purification, partial characterization and biological effects of the XTC mesoderm-inducing factor. 1989, Pubmed , Xenbase
Smith, Mesoderm induction and the control of gastrulation in Xenopus laevis: the roles of fibronectin and integrins. 1990, Pubmed , Xenbase
Sumoy, PACSIN 3 is a novel SH3 domain cytoplasmic adapter protein of the pacsin-syndapin-FAP52 gene family. 2001, Pubmed
Tadokoro, Talin binding to integrin beta tails: a final common step in integrin activation. 2003, Pubmed
Tseng, The bimodal role of filamin in controlling the architecture and mechanics of F-actin networks. 2004, Pubmed
Turner, Expression of achaete-scute homolog 3 in Xenopus embryos converts ectodermal cells to a neural fate. 1994, Pubmed , Xenbase
Webb, Adhesion assembly, disassembly and turnover in migrating cells -- over and over and over again. 2002, Pubmed
Winklbauer, Fibronectin, mesoderm migration, and gastrulation in Xenopus. 1996, Pubmed , Xenbase
van der Flier, Structural and functional aspects of filamins. 2001, Pubmed
van der Flier, Different splice variants of filamin-B affect myogenesis, subcellular distribution, and determine binding to integrin [beta] subunits. 2002, Pubmed
Alfandari, Integrin alpha5beta1 supports the migration of Xenopus cranial neural crest on fibronectin. 2003, Pubmed , Xenbase
Alfandari, Integrin alpha v subunit is expressed on mesodermal cell surfaces during amphibian gastrulation. 1995, Pubmed , Xenbase
Alfandari, ADAM 13: a novel ADAM expressed in somitic mesoderm and neural crest cells during Xenopus laevis development. 1997, Pubmed , Xenbase
Badour, The Wiskott-Aldrich syndrome protein acts downstream of CD2 and the CD2AP and PSTPIP1 adaptors to promote formation of the immunological synapse. 2003, Pubmed
Boucaut, Prevention of gastrulation but not neurulation by antibodies to fibronectin in amphibian embryos. 1984, Pubmed
Boucaut, Biologically active synthetic peptides as probes of embryonic development: a competitive peptide inhibitor of fibronectin function inhibits gastrulation in amphibian embryos and neural crest cell migration in avian embryos. 1984, Pubmed
Bubeck, Ligand recruitment by vinculin domains in transfected cells. 1997, Pubmed
Calderwood, Talin controls integrin activation. 2004, Pubmed
Chitu, Pombe Cdc15 homology (PCH) proteins: coordinators of membrane-cytoskeletal interactions. 2007, Pubmed
Cohen, Active zones on motor nerve terminals contain alpha 3beta 1 integrin. 2000, Pubmed , Xenbase
Cousin, PACSIN2 is a regulator of the metalloprotease/disintegrin ADAM13. 2000, Pubmed , Xenbase
Cousin, A PTP-PEST-like protein affects alpha5beta1-integrin-dependent matrix assembly, cell adhesion, and migration in Xenopus gastrula. 2004, Pubmed , Xenbase
Cunningham, Actin-binding protein requirement for cortical stability and efficient locomotion. 1992, Pubmed
Darribère, Distinct regions of human fibronectin are essential for fibril assembly in an in vivo developing system. 1992, Pubmed
Darribère, In vivo analyses of integrin beta 1 subunit function in fibronectin matrix assembly. 1990, Pubmed
Davidson, Mesendoderm extension and mantle closure in Xenopus laevis gastrulation: combined roles for integrin alpha(5)beta(1), fibronectin, and tissue geometry. 2002, Pubmed , Xenbase
Davidson, Integrin alpha5beta1 and fibronectin regulate polarized cell protrusions required for Xenopus convergence and extension. 2006, Pubmed , Xenbase
Fankhauser, The S. pombe cdc15 gene is a key element in the reorganization of F-actin at mitosis. 1995, Pubmed
Flanagan, Filamin A, the Arp2/3 complex, and the morphology and function of cortical actin filaments in human melanoma cells. 2001, Pubmed
Gaultier, ADAM13 disintegrin and cysteine-rich domains bind to the second heparin-binding domain of fibronectin. 2002, Pubmed , Xenbase
Gawantka, A beta 1-integrin associated alpha-chain is differentially expressed during Xenopus embryogenesis. 1995, Pubmed , Xenbase
Ghadimi, Identification of interaction partners of the cytosolic polyproline region of CD95 ligand (CD178). 2002, Pubmed
Ginsberg, Integrin regulation. 2005, Pubmed
Glogauer, The role of actin-binding protein 280 in integrin-dependent mechanoprotection. 1998, Pubmed
Halbach, PACSIN 1 forms tetramers via its N-terminal F-BAR domain. 2007, Pubmed
Hynes, Integrins: bidirectional, allosteric signaling machines. 2002, Pubmed
Jha, Evidence for the involvement of proline-directed serine/threonine phosphorylation in sperm capacitation. 2006, Pubmed
Joos, Integrin alpha 5 during early development of Xenopus laevis. 1995, Pubmed , Xenbase
Keller, Mechanisms of convergence and extension by cell intercalation. 2000, Pubmed
Kessels, The syndapin protein family: linking membrane trafficking with the cytoskeleton. 2004, Pubmed
Kessels, Syndapin oligomers interconnect the machineries for endocytic vesicle formation and actin polymerization. 2006, Pubmed
Kiema, The molecular basis of filamin binding to integrins and competition with talin. 2006, Pubmed
Lee, Temporal and spatial regulation of fibronectin in early Xenopus development. 1984, Pubmed , Xenbase
Li, A cdc15-like adaptor protein (CD2BP1) interacts with the CD2 cytoplasmic domain and regulates CD2-triggered adhesion. 1999, Pubmed
Marsden, Regulation of cell polarity, radial intercalation and epiboly in Xenopus: novel roles for integrin and fibronectin. 2001, Pubmed , Xenbase
Marsden, Integrin-ECM interactions regulate cadherin-dependent cell adhesion and are required for convergent extension in Xenopus. 2003, Pubmed , Xenbase
Modregger, All three PACSIN isoforms bind to endocytic proteins and inhibit endocytosis. 2001, Pubmed
Mori, PACSIN3 binds ADAM12/meltrin alpha and up-regulates ectodomain shedding of heparin-binding epidermal growth factor-like growth factor. 2003, Pubmed
Mould, Regulation of integrin alpha 5 beta 1-fibronectin interactions by divalent cations. Evidence for distinct classes of binding sites for Mn2+, Mg2+, and Ca2+. 1995, Pubmed
Na, Differential regulation of cell adhesive functions by integrin alpha subunit cytoplasmic tails in vivo. 2003, Pubmed , Xenbase
Nikki, FAP52 regulates actin organization via binding to filamin. 2002, Pubmed
Nikki, Focal adhesion protein FAP52 self-associates through a sequence conserved among the members of the PCH family proteins. 2002, Pubmed
Park, Subcellular localization and signaling properties of dishevelled in developing vertebrate embryos. 2005, Pubmed , Xenbase
Pistor, The ActA protein of Listeria monocytogenes acts as a nucleator inducing reorganization of the actin cytoskeleton. 1994, Pubmed
Qualmann, Syndapin isoforms participate in receptor-mediated endocytosis and actin organization. 2000, Pubmed
Ramos, Xenopus embryonic cell adhesion to fibronectin: position-specific activation of RGD/synergy site-dependent migratory behavior at gastrulation. 1996, Pubmed , Xenbase
Ramos, Integrin-dependent adhesive activity is spatially controlled by inductive signals at gastrulation. 1996, Pubmed , Xenbase
Ridley, Cell migration: integrating signals from front to back. 2003, Pubmed
Simpson, SH3-domain-containing proteins function at distinct steps in clathrin-coated vesicle formation. 1999, Pubmed
Smith, Purification, partial characterization and biological effects of the XTC mesoderm-inducing factor. 1989, Pubmed , Xenbase
Smith, Mesoderm induction and the control of gastrulation in Xenopus laevis: the roles of fibronectin and integrins. 1990, Pubmed , Xenbase
Sumoy, PACSIN 3 is a novel SH3 domain cytoplasmic adapter protein of the pacsin-syndapin-FAP52 gene family. 2001, Pubmed
Tadokoro, Talin binding to integrin beta tails: a final common step in integrin activation. 2003, Pubmed
Tseng, The bimodal role of filamin in controlling the architecture and mechanics of F-actin networks. 2004, Pubmed
Turner, Expression of achaete-scute homolog 3 in Xenopus embryos converts ectodermal cells to a neural fate. 1994, Pubmed , Xenbase
Webb, Adhesion assembly, disassembly and turnover in migrating cells -- over and over and over again. 2002, Pubmed
Winklbauer, Fibronectin, mesoderm migration, and gastrulation in Xenopus. 1996, Pubmed , Xenbase
van der Flier, Structural and functional aspects of filamins. 2001, Pubmed
van der Flier, Different splice variants of filamin-B affect myogenesis, subcellular distribution, and determine binding to integrin [beta] subunits. 2002, Pubmed