SH3-dependent stimulation of Src-family kinase autophosphorylation without tail release from the SH2 domain in vivo

Lerner, Edwina C.; Smithgall, Thomas E.

doi:10.1038/nsb782

Cited by 84 publications

(175 citation statements)

References 29 publications

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“…4b). Similarly to our finding, Lerner and Smithgall also showed that human immunodeficiency virus-1 Nef proteininduced Hck (another Src family member) activation is not associated with a decrease in the phosphorylation of the conserved C-terminal tyrosine (35). It is possible that SH3 and/or SH2 domain ligand-induced activation of Src family protein tyrosine kinases is a direct effect and does not require the activation of protein tyrosine phosphatases.…”

Section: Mechanical Stretch-induced C-src Activation and C-srcsupporting

confidence: 61%

Conversion of Mechanical Force into Biochemical Signaling

Han

Bai

Lodyga

et al. 2004

Journal of Biological Chemistry

141

132

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Physical forces play important roles in regulating cell proliferation, differentiation, and death by activating intracellular signal transduction pathways. How cells sense mechanical stimulation, however, is largely unknown. Most studies focus on cellular membrane proteins such as ion channels, integrins, and receptors for growth factors as mechanosensory units. Here we show that mechanical stretch-induced c-Src protein tyrosine kinase activation is mediated through the actin filament-associated protein (AFAP). Distributed along the actin filaments, AFAP can directly active c-Src through binding to its Src homology 3 and/or 2 domains. Mutations at these specific binding sites on AFAP blocked mechanical stretch-induced c-Src activation. Therefore, mechanical force can be transmitted along the cytoskeleton, and interaction between cytoskeletal associated proteins and enzymes related to signal transduction may convert physical forces into biochemical reactions. Cytoskeleton deformation-induced proteinprotein interaction via specific binding sites may represent a novel intracellular mechanism for cells to sense mechanical stimulation.

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Section: Mechanical Stretch-induced C-src Activation and C-srcsupporting

confidence: 61%

Conversion of Mechanical Force into Biochemical Signaling

Han

Bai

Lodyga

et al. 2004

Journal of Biological Chemistry

141

132

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“…S6). A number of in vitro and in vivo data including cell based assays and mutagenesis have also supported the concept of such an SH3-displaced model in kinase activation (5,27). While the present results are clearly at the limit of resolution available from the analysis of SAXS data, they support the concept of SH3-displaced states.…”

Section: Discussioncontrasting

confidence: 46%

Multidomain assembled states of Hck tyrosine kinase in solution

Yang

Blachowicz

Makowski

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

193

199

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An approach combining small-angle X-ray solution scattering (SAXS) data with coarse-grained (CG) simulations is developed to characterize the assembly states of Hck, a member of the Srcfamily kinases, under various conditions in solution. First, a basis set comprising a small number of assembly states is generated from extensive CG simulations. Second, a theoretical SAXS profile for each state in the basis set is computed by using the Fast-SAXS method. Finally, the relative population of the different assembly states is determined via a Bayesian-based Monte Carlo procedure seeking to optimize the theoretical scattering profiles against experimental SAXS data. The study establishes the concept of basis-set supported SAXS (BSS-SAXS) reconstruction combining computational and experimental techniques. Here, BSS-SAXS reconstruction is used to reveal the structural organization of Hck in solution and the different shifts in the equilibrium population of assembly states upon the binding of different signaling peptides.T yrosine kinases of the Src-family are large multidomain allosteric enzymes implicated in the signaling pathways regulating cell growth and proliferation (1). The key role that Src kinases play in the onset of many human diseases, particularly cancer, makes them important targets for therapeutic intervention (2).All Src kinases share a common structural organization comprising the SH3 and SH2 binding domains followed by a highly conserved catalytic domain connected by flexible linkers (1). Crystal structures have offered a detailed view of the down-regulated forms of Hck and c-Src (3, 4), characterized by a compact assembled form stabilized by auto-inhibitory intramolecular interactions between the N and C terminus of the catalytic domain with the SH3 and SH2 modules, respectively. The SH2 and SH3 modules of the Src-family tyrosine kinases play dual roles: both are involved in the auto-inhibitory intramolecular interactions down-regulating kinase activity, but can also serve as possible binding receptors for cellular signals leading to kinase activation.In a broadly accepted paradigm, Src inactivation/activation is pictured in terms of a simple two-state process involving an assembled (inactive) and a disassembled (active) conformation, i.e., once any of the intramolecular interaction is released, the kinase domain switches to a disassembled catalytically active state targeting down-stream cellular substrates (5-8). Crystal structures (3, 4) and mutational studies complemented by molecular dynamics (MD) simulations (9) are consistent with this view. However, one crystal structure of c-Src in a partially activated state in which the SH2-SH3 modules are reassembled in a different orientation with respect to the catalytic domain (10), suggests that the situation could be considerably more complex. In this regard, it seems likely that different signals could promote different assembly states, leading to differently configured activated kinases. While it is understood that Src increases its activity in respon...

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“…Then what is the role of C3G in the induction of apoptosis? Activation of Hck by mutation of negative regulatory tyrosine phosphorylation site or by the SH3 ligand Nef results in fibroblast cell transformation (45)(46)(47). This is due to large increases in kinase activity of Hck.…”

Section: Discussionmentioning

confidence: 99%

Physical and Functional Interaction between Hck Tyrosine Kinase and Guanine Nucleotide Exchange Factor C3G Results in Apoptosis, Which Is Independent of C3G Catalytic Domain

Shivakrupa

Radha

Sudhakar

et al. 2003

Journal of Biological Chemistry

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The hematopoietic cell kinase Hck is a Src family tyrosine kinase expressed in cells of myelomonocytic lineage, B lymphocytes, and embryonic stem cells. To study its role in signaling pathways we used the Hck-SH3 domain in protein interaction cloning and identified C3G, the guanine nucleotide exchange factor for Rap1 and R-Ras, as a protein that associated with Hck. This interaction was direct and was mediated partly through the proline-rich region of C3G. C3G could be co-immunoprecipitated with Hck from Cos-1 cells transfected with Hck and C3G. C3G was phosphorylated on tyrosine 504 in cells when coexpressed with Hck but not with a catalytically inactive mutant of Hck. Phosphorylation of endogenous C3G at Tyr-504 was increased by treatment of human myelomonocytic THP-1 cells with mercuric chloride, which is known to activate Hck tyrosine kinase specifically. Coexpression of Hck with C3G induced a high level of apoptosis in many cell lines by 30 -42 h of transfection. Induction of apoptosis was not dependent on Tyr-504 phosphorylation or the catalytic domain of C3G but required the catalytic activity of Hck. Using dominant negative constructs of caspases we found that caspase-1, -8, and -9 are involved in this apoptotic pathway. These results suggest that C3G and Hck interact physically and functionally in vivo to activate kinase-dependent and caspase-mediated apoptosis, which is independent of catalytic domain of C3G.The Src family tyrosine kinases play an important role in linking signals received by transmembrane receptors and a variety of intracellular pathways, thereby regulating diverse cellular responses such as proliferation, differentiation, and cell death (1, 2). This is achieved through their non-catalytic sequences, which enable multiple interactions with cellular proteins, and through the kinase domain, which phosphorylates substrates to alter their activity, change their subcellular location, and effect their intermolecular interactions. The hematopoietic cell kinase (Hck) 1 is a Src family member that is expressed in cells of myelomonocytic lineage, B lymphocytes, and embryonic stem cells with higher levels in differentiated cells, suggesting a role for this enzyme in signaling pathways of mature hematopoietic cells (3-5). Hck is activated by agents that induce macrophage differentiation and in response to cytokines such as interleukin-3, granulocyte-macrophage colony stimulating factor, and leukemia inhibitory factor. It is also involved in cytokine production in macrophages in response to lipopolysaccharide and viral infection (6 -9).Structurally, Hck is similar to other members of the Src family in that it has a catalytic domain at the C terminus that is preceded by a 100-amino acid SH2 domain and a 50-amino acid SH3 domain. The SH2 and SH3 domains are protein interaction modules that mediate either intramolecular or intermolecular associations. SH2 domains bind to phosphorylated tyrosine residues in a specific amino acid context, whereas SH3 domain interacts with polyproline tracts in polypept...

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SH3-dependent stimulation of Src-family kinase autophosphorylation without tail release from the SH2 domain in vivo

Cited by 84 publications

References 29 publications

Conversion of Mechanical Force into Biochemical Signaling

Conversion of Mechanical Force into Biochemical Signaling

Multidomain assembled states of Hck tyrosine kinase in solution

Physical and Functional Interaction between Hck Tyrosine Kinase and Guanine Nucleotide Exchange Factor C3G Results in Apoptosis, Which Is Independent of C3G Catalytic Domain

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