Multiple Domains of Ruk/CIN85/SETA/CD2BP3 are Involved in Interaction with p85α Regulatory Subunit of PI 3-kinase

Borthwick, Emma B.; Коробко, И. В.; Luke, Courtney T.; Drel, Viktor R.; Fedyshyn, Yaroslav; Ninkina, Natalia; Дробот, Л. Б.; Buchman, Vladimir L.

doi:10.1016/j.jmb.2004.08.075

Cited by 24 publications

(32 citation statements)

References 32 publications

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“…When the number of unique peptides identified using the bait was not at least 5 times greater compared with the control, proteins were excluded from the hit list. Through this approach we could identify 50 proteins, which confirm CIN85 itself, Cbl, Alix, ARAP1, Dynamin-2, BLNK, and SHIP-1 as binding partners of CIN85 as known from other cellular systems, for example (14,17,(32)(33)(34)(35). Moreover, Net1, BCAP, HPK1, Rhotekin-2, PP1␣, and CHTF8 were identified as novel interaction partners for CIN85's SH3 domains (Table I).…”

Section: Cin85 Interacts With Ship-1 In B Cellsmentioning

confidence: 82%

“…We therefore postulated that the SH3 domains of other proteins may displace CIN85's SH3 domains. This in turn would allow the SH3 domains of CIN85 to interact with other proteins, as shown for the p85 subunit of the PI-3-kinase (32). Therefore, it would be conceivable that a broader spectrum of proteins could interact with the free SH3 domains of CIN85, compared with the dimerized full-length molecule.…”

Section: Cin85 Interacts With Ship-1 In B Cellsmentioning

confidence: 99%

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CIN85 Interacting Proteins in B Cells-Specific Role for SHIP-1

Büchse

Horras

Lenfert

et al. 2011

Molecular & Cellular Proteomics

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The Cbl-interacting 85-kDa protein (CIN85) plays an important role as a negative regulator of signaling pathways induced by receptor tyrosine kinases. By assembling multiprotein complexes this versatile adaptor enhances receptor tyrosine kinase-activated clathrin-mediated endocytosis and reduces phosphatidylinositol-3-kinase-induced phosphatidylinositol-3,4,5-trisphosphate production. Here we report the expression of CIN85 in primary splenic B lymphocytes and the B-lymphoma cell lines WEHI 231 and Ba/F3. Cross-linking of the B cell antigen receptor resulted in an increased association of CIN85 with the ubiquitin ligase Cbl. Through a systematic pull-down proteomics approach we identified 51 proteins that interact with CIN85 in B cells, including proteins not shown previously to be CIN85-associated. Among these proteins, the SH2-containing inositol phosphatase 1 (SHIP-1) co-precipitated with both the fulllength CIN85 and each of its three SH3 domains. We also showed that this association is constitutive and depends on a region of 79 amino acids near the carboxyl terminus of SHIP-1, a region rich in potential SH3 domain binding sites. Because SHIP-1 is a major negative regulator of the phosphatidylinositol-3-kinase pathway in lymphocytes, we hypothesize that the interaction between SHIP-1 and CIN85 might synergistically facilitate the down-regulation of phosphatidylinositol-3,4,5-trisphosphate levels. Molecular &

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Section: Cin85 Interacts With Ship-1 In B Cellsmentioning

confidence: 82%

Section: Cin85 Interacts With Ship-1 In B Cellsmentioning

confidence: 99%

CIN85 Interacting Proteins in B Cells-Specific Role for SHIP-1

Büchse

Horras

Lenfert

et al. 2011

Molecular & Cellular Proteomics

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“…In some experiments, FLAG-tagged CIN85 mutants lacking distinct domains or alanine mutants in the proline-rich region of CIN85 were used as described before (21). Cells transfected with empty FLAG and HA vectors (EV), HA-tagged dual-specificity phosphatase 7 (DUSP7) (gift from Igor Astsaturov; Addgene plasmid #27976), and FLAG-tagged CIN85 served as controls.…”

Section: Methodsmentioning

confidence: 99%

“…To further map the PP2Ac binding domain within the Pro-rich region, we utilized the full-length CIN85 plasmids wherein several proline residues or adjacent charged amino acids within one of each Pro-rich blocks (P1 through P4) were substituted with alanine (21) (Fig. 2C).…”

Section: Cin85 Is a Novel Pp2acmentioning

confidence: 99%

A Novel Interaction of the Catalytic Subunit of Protein Phosphatase 2A with the Adaptor Protein CIN85 Suppresses Phosphatase Activity and Facilitates Platelet Outside-in αIIbβ3 Integrin Signaling

Khatlani¹,

Pradhan²,

Da³

et al. 2016

Journal of Biological Chemistry

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The transduction of signals generated by protein kinases and phosphatases are critical for the ability of integrin ␣ IIb ␤ 3 to support stable platelet adhesion and thrombus formation. Unlike kinases, it remains unclear how serine/threonine phosphatases engage the signaling networks that are initiated following integrin ligation. Because protein-protein interactions form the backbone of signal transduction, we searched for proteins that interact with the catalytic subunit of protein phosphatase 2A (PP2Ac). In a yeast two-hybrid study, we identified a novel interaction between PP2Ac and an adaptor protein CIN85 (Cbl-interacting protein of 85 kDa). Truncation and alanine mutagenesis studies revealed that PP2Ac binds to the P3 block ( 396 PAIPPKKPRP 405 ) of the proline-rich region in CIN85. The interaction of purified PP2Ac with CIN85 suppressed phosphatase activity. Human embryonal kidney 293 ␣ IIb ␤ 3 cells overexpressing a CIN85 P3 mutant, which cannot support PP2Ac binding, displayed decreased adhesion to immobilized fibrinogen. Platelets contain the ϳ85 kDa CIN85 protein along with the PP2Ac-CIN85 complex. A myristylated cell-permeable peptide derived from residues 395-407 of CIN85 protein (P3 peptide) disrupted the platelet PP2Ac-CIN85 complex and decreased ␣ IIb ␤ 3 signaling dependent functions such as platelet spreading on fibrinogen and thrombin-mediated fibrin clot retraction. In a phospho-profiling study P3 peptide treated platelets also displayed decreased phosphorylation of several signaling proteins including Src and GSK3␤. Taken together, these data support a role for the novel PP2Ac-CIN85 complex in supporting integrindependent platelet function by dampening the phosphatase activity.An ischemic vascular event ensues when a blood vessel is occluded by platelet-rich thrombi at the site of ruptured atherosclerotic plaque. Exposure of platelets to collagen and/or von Willebrand factor under high shear stress within the milieu of plaque initiates the inside-out signaling processes that activates integrin ␣ IIb ␤ 3 (1). The activated integrin can engage fibrinogen and generate outside-in signals that facilitate cytoskeletal reorganization and support a cascade of functional changes, including platelet spreading, stable adhesion, granule secretion, and clot retraction; all processes that augment the size of the platelet thrombi (2, 3).The coordinated signaling networks that are initiated by the engagement of agonists and integrin receptors on platelets are essential to the development of thrombi (4, 5). Spatial and temporal assembly of multiprotein signaling complexes by adaptor proteins relay signals following the engagement of integrin. The reversible phosphorylation of proteins on tyrosine, serine, and threonine residues, under the control of kinases and phosphatases, is an essential component of signaling. Because the outside-in signaling process greatly amplifies the size of platelet thrombi, there is a desire to selectively target this signaling process in a new class of antithrombotic agents (6). ...

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“…Overexpression of Ruk l /CIN85 full-length form induces apoptotic cell death of primary neurons in culture [2,15]. However, shorter molecular forms of Ruk/CIN85 block the pro-apoptotic effect of Ruk l /CIN85, suggesting that expression of different combinations of Ruk/CIN85 proteins in cells could be involved in the regulation of their survival and other intracellular processes [2,15]. CD2AP/CMS is required for rapid activation of PI3K and ERK/MAPK pathways by TGF-β [29].…”

Section: Introductionmentioning

confidence: 99%

Comparative study of expression of adaptor proteins Ruk/CIN85 and CD2AP/CMS in normal andtumor human uterus tissues

et al. 2009

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Adaptor proteins play an important role in facilitating protein-protein interactions and subsequent formation of signalling networks. These proteins recruit binding partners to a specific location inside the cell, and also regulate their activity. Adaptor protein Ruk/CIN85 and its structural and functional homologue CD2AP/CMS are important components of different regulatory pathways involved in control of cell proliferation, adhesion, invasion and survival, and, thus, can play a role in uterine carcinogenesis. In this work, we set out a comparative study of expression of Ruk/CIN85 and CD2AP/CMS at the level of mRNA and protein in intact uterine tissues, as well as in benign and malignant uterine tumors of different histological types. In most cases, an increase of expression levels of Ruk/CIN85 full-length form mRNA and protein, as well as CD2AP/CMS protein, were observed in uterine tumors, comparing with surrounding normal uterine tissues. Characteristic feature of conditionally normal uterine tissues, as well as benign uterine lesions, was an elevated content of high-molecular mass Ruk/CIN85 forms of 140 and 130 kDa, while an increased expression level of low-molecular mass 40 and 30 kDa Ruk/CIN85 forms was observed in the malignant tissue samples. Our findings suggest that an abnormal expression patterns of adaptor proteins Ruk/CIN85 and CD2AP/CMS in uterine tumors, and, thus, the corresponding changes in the activity of downstream signalling pathways, might be involved in maintenance of the malignant phenotype.Key words: tumorigenesis, adaptor proteins, Ruk/CIN85, CD2AP/CMS, benign and malignant uterine tumors.Abbreviations: Ruk/CIN85, regulator of ubiquitous kinase/Cbl-interacting protein of 85 kDa; CD2AP/CMS, CD2 associated protein/Cas ligand with multiple SH3 domains; Biol. Stud. 2009: 3(3); 5-16 •

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Multiple Domains of Ruk/CIN85/SETA/CD2BP3 are Involved in Interaction with p85α Regulatory Subunit of PI 3-kinase

Cited by 24 publications

References 32 publications

CIN85 Interacting Proteins in B Cells-Specific Role for SHIP-1

CIN85 Interacting Proteins in B Cells-Specific Role for SHIP-1

A Novel Interaction of the Catalytic Subunit of Protein Phosphatase 2A with the Adaptor Protein CIN85 Suppresses Phosphatase Activity and Facilitates Platelet Outside-in αIIbβ3 Integrin Signaling

Comparative study of expression of adaptor proteins Ruk/CIN85 and CD2AP/CMS in normal andtumor human uterus tissues

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