Molecular Cloning of p125Nap1, a Protein That Associates with an SH3 Domain of Nck

Kitamura, Tadahiro; Kitamura, Yoichi; Yonezawa, Kazuyoshi; Nf, Totty; Gout, Ivan; Hara, Kenta; Waterfield,; Sakaue, Motoyoshi; Ogawa, Wataru; Kasuga, Masato

doi:10.1006/bbrc.1996.0264

Cited by 62 publications

(53 citation statements)

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“…Polyclonal anti-Nap1 antiserum was raised against a C-terminal peptide (CHAVYKQSVTSSA), as described (Eden et al, 2002;Kitamura et al, 1996;Steffen et al, 2004). Anti-Abi1 monoclonal antibody was a generous gift from G. Scita (Milan).…”

Section: Antibodies and Reagentsmentioning

confidence: 99%

Axis specification and morphogenesis in the mouse embryo requireNap1, a regulator of WAVE-mediated actin branching

2006

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Dynamic cell movements and rearrangements are essential for the generation of the mammalian body plan, although relatively little is known about the genes that coordinate cell movement and cell fate. WAVE complexes are regulators of the actin cytoskeleton that couple extracellular signals to polarized cell movement. Here, we show that mouse embryos that lack Nap1, a regulatory component of the WAVE complex, arrest at midgestation and have defects in morphogenesis of all three embryonic germ layers. WAVE protein is not detectable in Nap1 mutants, and other components of the WAVE complex fail to localize to the surface of Nap1 mutant cells; thus loss of Nap1 appears to inactivate the WAVE complex in vivo. Nap1 mutants show specific morphogenetic defects: they fail to close the neural tube, fail to form a single heart tube (cardia bifida), and show delayed migration of endoderm and mesoderm. Other morphogenetic processes appear to proceed normally in the absence of Nap1/WAVE activity: the notochord, the layers of the heart, and the epithelial-to-mesenchymal transition (EMT) at gastrulation appear normal. A striking phenotype seen in approximately one quarter of Nap1 mutants is the duplication of the anteroposterior body axis. The axis duplications arise because Nap1 is required for the normal polarization and migration of cells of the Anterior Visceral Endoderm (AVE), an early extraembryonic organizer tissue. Thus, the Nap1 mutant phenotypes define the crucial roles of Nap1/WAVE-mediated actin regulation in tissue organization and establishment of the body plan of the mammalian embryo.

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Section: Antibodies and Reagentsmentioning

confidence: 99%

Axis specification and morphogenesis in the mouse embryo requireNap1, a regulator of WAVE-mediated actin branching

2006

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“…The 125 kDa NAP1 (Nck-associated protein 1) is a receptor-like protein with six transmembrane domains that belongs to the evolutionarily-conserved Hem gene family (Kitamura et al, 1996). NAP1 indirectly associates, through a 140 kDa protein, with the GTPbound Rac1 (Kitamura et al, 1997), and binds directly to the N-terminal SH3 domain of Nck (Kitamura et al, 1996).…”

Section: Nck-nap1 (Hem-2 or Kette) Complex In Axon Pathfinding And Mementioning

confidence: 99%

“…NAP1 indirectly associates, through a 140 kDa protein, with the GTPbound Rac1 (Kitamura et al, 1997), and binds directly to the N-terminal SH3 domain of Nck (Kitamura et al, 1996). While the function of Hem proteins in mammals remains unclear, its Drosophila homologue Kette/Hem-2 appears to act either as a cell surface receptor or a membrane docking protein to control VUM axon path®nding and cytoskeletal organization (Hummel et al, 2000).…”

Section: Nck-nap1 (Hem-2 or Kette) Complex In Axon Pathfinding And Mementioning

confidence: 99%

Nck/Dock: an adapter between cell surface receptors and the actin cytoskeleton

2001

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In response to extracellular signals, cell surface receptors engage in connections with multiple intracellular signaling pathways, leading to the cellular responses such as survival, migration, proliferation and di erentiation. Thè pY?SH2/SH3?e ector' connection is a frequently used scheme by many cell surface receptors, in which SH2/SH3-containing adapters connect protein tyrosine phosphorylation to a variety of downstream e ector pathways. Following the initial landmark ®nding that Grb2 adapter links the receptors to the Ras pathway leading to DNA synthesis, recent studies have revealed that the biological function of the SH2/SH3 adapter Nck/Dock is to link cell surface receptors to the actin cytoskeleton. For example, in the evolutionarily-conserved signaling network, GEF-Rac-Nck-Pak, Nck`®xes up' the interaction of Pak with its upstream activator, Rac. The activated Pak then regulates the cytoskeletal dynamics. The fact that the majority of the more than 20 Nck-SH3-associated e ectors are regulators of the actin cytoskeleton suggests that Nck/Dock regulates, via binding to distinct e ectors, various cell type-speci®c motogenic responses. This review focuses on our current understanding of Nck/Dock function. Due to the number and complexity of the terminologies used in this review, à Glossary of Terms' is provided to help reduce confusions. Oncogene (2001) 20, 6403 ± 6417.Keywords: Src-homology domains; adapters; tyrosine kinases; signal transduction; cytoskeleton The SH2/SH3 adapters The src-homology 2 (SH2) and src-homlogy 3 (SH3) domains are protein ± protein interaction modules, found in a variety of structurally and functionally distinct signaling molecules, including protein tyrosine kinases/phosphatases, lipid kinase/phosphatase, guanine nucleotide exchange factors/GTPase activation proteins, docking proteins/adapters, cytoskeletal binding proteins and even transcription factors (Pawson, 1995;Mayer, 1999). Except for the terminology, SH2 and SH3 domains share no other common features regarding their structures, binding speci®city, and cellular targets. SH2 domains enable their host proteins to form complexes with phosphotyrosine (pY) proteins by binding to pY residues within these proteins. The stability and speci®city of the binding also depend upon the additional three to six amino acid residues immediately carboxyl to the pY residue, pY-(X) 3 ± 6 . Therefore, SH2 domains from di erent host molecules bind di erent pY-containing peptides. SH3 domains bind proline-rich segments of target molecules with a minimum consensus of P-x-x-P (Mayer, 2001). Each proline is often preceded by an aliphatic amino acid residue, and the aliphatic-proline pair binds to a hydrophobic pocket on SH3 domains. A major di erence between the SH2-pY binding and SH3-PxxP interactions is that the former is always transiently induced by extracellular signals, whereas the latter is often, but not always, constitutive. For instance, the SH2 domain of the adapter protein Grb2 only binds to tyrosine phosphorylated/activated EGFR, whe...

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“…Lee et al, 1993;Chou and Hanafusa, 1995;Hu et al, 1995;Rivero-Lezcano et al, 1995;Birge et al, 1996;Bokoch et al, 1996;Galisteo et al, 1996;Kitamura et al, 1996;Quilliam et al, 1996;Lu et al, 1997;Lussier and Larose, 1997;Su et al, 1997;Anton et al, 1998). Nck is capable of physically associating with the signaling proteins through direct or indirect protein-protein interactions.…”

Section: Introductionmentioning

confidence: 99%

Nck-2, a Novel Src Homology2/3-containing Adaptor Protein That Interacts with the LIM-only Protein PINCH and Components of Growth Factor Receptor Kinase-signaling Pathways

1998

MBoC

167

166

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Many of the protein-protein interactions that are essential for eukaryotic intracellular signal transduction are mediated by protein binding modules including SH2, SH3, and LIM domains. Nck is a SH3-and SH2-containing adaptor protein implicated in coordinating various signaling pathways, including those of growth factor receptors and cell adhesion receptors. We report here the identification, cloning, and characterization of a widely expressed, Nck-related adaptor protein termed Nck-2. Nck-2 comprises primarily three Nterminal SH3 domains and one C-terminal SH2 domain. We show that Nck-2 interacts with PINCH, a LIM-only protein implicated in integrin-linked kinase signaling. The PINCHNck-2 interaction is mediated by the fourth LIM domain of PINCH and the third SH3 domain of Nck-2. Furthermore, we show that Nck-2 is capable of recognizing several key components of growth factor receptor kinase-signaling pathways including EGF receptors, PDGF receptor-␤, and IRS-1. The association of Nck-2 with EGF receptors was regulated by EGF stimulation and involved largely the SH2 domain of Nck-2, although the SH3 domains of Nck-2 also contributed to the complex formation. The association of Nck-2 with PDGF receptor-␤ was dependent on PDGF activation and was mediated solely by the SH2 domain of Nck-2. Additionally, we have detected a stable association between Nck-2 and IRS-1 that was mediated primarily via the second and third SH3 domain of Nck-2. Thus, Nck-2 associates with PINCH and components of different growth factor receptor-signaling pathways via distinct mechanisms. Finally, we provide evidence indicating that a fraction of the Nck-2 and/or Nck-1 proteins are associated with the cytoskeleton. These results identify a novel Nck-related SH2-and SH3-domain-containing protein and suggest that it may function as an adaptor protein connecting the growth factor receptor-signaling pathways with the integrin-signaling pathways.

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Molecular Cloning of p125Nap1, a Protein That Associates with an SH3 Domain of Nck

Cited by 62 publications

References 0 publications

Axis specification and morphogenesis in the mouse embryo requireNap1, a regulator of WAVE-mediated actin branching

Axis specification and morphogenesis in the mouse embryo requireNap1, a regulator of WAVE-mediated actin branching

Nck/Dock: an adapter between cell surface receptors and the actin cytoskeleton

Nck-2, a Novel Src Homology2/3-containing Adaptor Protein That Interacts with the LIM-only Protein PINCH and Components of Growth Factor Receptor Kinase-signaling Pathways

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