All Title Author
Keywords Abstract

PLOS ONE  2013 

Adaptor Proteins Intersectin 1 and 2 Bind Similar Proline-Rich Ligands but Are Differentially Recognized by SH2 Domain-Containing Proteins

DOI: 10.1371/journal.pone.0070546

Full-Text   Cite this paper   Add to My Lib

Abstract:

Background Scaffolding proteins of the intersectin (ITSN) family, ITSN1 and ITSN2, are crucial for the initiation stage of clathrin-mediated endocytosis. These proteins are closely related but have implications in distinct pathologies. To determine how these proteins could be separated in certain cell pathways we performed a comparative study of ITSNs. Methodology/Principal Findings We have shown that endogenous ITSN1 and ITSN2 colocalize and form a complex in cells. A structural comparison of five SH3 domains, which mediated most ITSNs protein-protein interactions, demonstrated a similarity of their ligand-binding sites. We showed that the SH3 domains of ITSN2 bound well-established interactors of ITSN1 as well as newly identified ITSNs protein partners. A search for a novel interacting interface revealed multiple tyrosines that could be phosphorylated in ITSN2. Phosphorylation of ITSN2 isoforms but not ITSN1 short isoform was observed in various cell lines. EGF stimulation of HeLa cells enhanced tyrosine phosphorylation of ITSN2 isoforms and enabled their recognition by the SH2 domains of the Fyn, Fgr and Abl1 kinases, the regulatory subunit of PI3K, the adaptor proteins Grb2 and Crk, and phospholipase C gamma. The SH2 domains mentioned were unable to bind ITSN1 short isoform. Conclusions/Significance Our results indicate that during evolution of vertebrates ITSN2 acquired a novel protein-interaction interface that allows its specific recognition by the SH2 domains of signaling proteins. We propose that these data could be important to understand the functional diversity of paralogous ITSN proteins.

References

[1]  Dacks JB, Field MC (2007) Evolution of the eukaryotic membrane-trafficking system: origin, tempo and mode. J Cell Sci 120: 2977–2985.
[2]  Henne MW, Boucrot E, Meinecke M, Evergren E, Vallis Y, et al. (2010) FCHo proteins are nucleators of clathrin-mediated endocytosis. Sci 328: 1281–1284.
[3]  Sengar AS, Wang W, Bishay J, Cohen S, Egan SE (1999) The EH and SH3 domain Ese proteins regulate endocytosis by linking to dynamin and Eps15. EMBO J 18: 1159–1171.
[4]  Pucharcos C, Casas C, Nadal M, Estivill X, de la Luna S (2001) The human intersectin genes and their spliced variants are differentially expressed. Biochim Biophys Acta 1521: 1–11.
[5]  Tsyba L, Nikolaienko O, Dergai O, Dergai M, Novokhatska O, et al. (2011) Intersectin multidomain adaptor proteins: Regulation of functional diversity. Gene 473: 67–75.
[6]  O’Bryan JP (2010) INTERSECTINg Pathways in Cell Biology. Sci Signal 3: re10.
[7]  Cataldo AM, Peterhoff CM, Troncoso JC, Gomez-Isla T, Hyman BT, et al. (2000) Endocytic pathway abnormalities precede amyloid beta deposition in sporadic Alzheimer’s disease and Down syndrome: differential effects of APOE genotype and presenilin mutations. Am J Pathol 157: 277–286.
[8]  Cataldo AM, Petanceska S, Peterhoff CM, Terio NB, Epstein CJ, et al. (2003) App gene dosage modulates endosomal abnormalities of Alzheimer’s disease in a segmental trisomy 16 mouse model of down syndrome. J Neurosci 23: 6788–6792.
[9]  Pucharcos C, Fuentes JJ, Casas C, de la Luna S, Alcantara S, et al. (1999) Alu-splice cloning of human Intersectin (ITSN), a putative multivalent binding protein expressed in proliferating and differentiating neurons and overexpressed in Down syndrome. Eur J Hum Genet 7: 704–712.
[10]  Fuentes JJ, Dierssen M, Pucharcos C, Fillat C, Casas C, et al. (1999) Application of Alu-splice PCR on chromosome 21: DSCR1 and Intersectin. J Neural Transm Suppl 57: 337–352.
[11]  Wilmot B, McWeeney SK, Nixon RR, Montine TJ, Laut J, et al. (2008) Translational gene mapping of cognitive decline. Neurobiol Aging 29: 524–541.
[12]  Hunter MP, Nelson M, Kurzer M, Wang X, Kryscio RJ, et al. (2011) Intersectin 1 contributes to phenotypes in vivo: implications for Down Syndrome. Neuroreport 22: 767–772.
[13]  Scappini E, Koh TW, Martin NP, O’Bryan JP (2007) Intersectin enhances huntingtin aggregation and neurodegeneration through activation of c-Jun-NH2-terminal kinase (JNK). Hum Mol Genet 16: 1862–1871.
[14]  Specht K, Harbeck N, Smida J, Annecke K, Reich U, et al. (2009) Expression profiling identifies genes that predict recurrence of breast cancer after adjuvant CMF-based chemotherapy. Breast Cancer Res Treat 118: 45–56.
[15]  McGavin M, Badour K, Hardy LA, Kubiseski TJ, Zhang J, et al. (2001) The intersectin 2 adaptor links Wiskott Aldrich syndrome protein (WASp)-mediated actin polymerization to T cell antigen receptor endocytosis. J Exp Med 194: 1777–1787.
[16]  Lim CS, Seet BT, Ingham RJ, Gish G, Matskova L, et al. (2007) The K15 protein of Kaposi’s sarcoma-associated herpesvirus recruits the endocytic regulator intersectin 2 through a selective SH3 domain interaction. Biochem 46: 9874–9885.
[17]  Asbach B, Ludwig C, Saksela K, Wagner R (2012) Comprehensive analysis of interactions between the Src-associated protein in mitosis of 68 kDa and the human Src-homology 3 proteome. PLoS ONE 7: e38540.
[18]  Klein I, Predescu D, Sharma T, Knezevic I, Malik A, et al. (2009) Intersectin-2L regulates caveola endocytosis secondary to Cdc42-mediated actin polymerization. J Biol Chem 284: 25953–25961.
[19]  Rodriguez-Fraticelli AE, Vergarajauregui S, Eastburn DJ, Datta A, Alonso MA, et al. (2010) The Cdc42 GEF Intersectin 2 controls mitotic spindle orientation to form the lumen during epithelial morphogenesis. J Cell Biol 189: 725–738.
[20]  Novokhatska O, Dergai M, Houssin N, Tsyba L, Moreau J, et al. (2011) Intersectin 2 nucleotide exchange factor regulates Cdc42 activity during Xenopus early development. Biochem Biophys Res Commun 408: 663–668.
[21]  Irie F, Yamaguchi Y (2002) EphB receptors regulate dendritic spine development via intersectin, Cdc42 and N-WASP. Nat Neurosci 5: 1117–1118.
[22]  Nishimura T, Yamaguchi T, Tokunaga A, Hara A, Hamaguchi T (2006) Role of Numb in dendritic spine development with a Cdc42 GEF intersectin and EphB2. Mol Biol Cell 17: 1273–1285.
[23]  Pechstein A, Shupliakov O, Haucke V (2010) Intersectin 1: a versatile actor in the synaptic vesicle cycle. Biochem Soc Trans 38: 181–186.
[24]  Tsyba L, Gryaznova T, Dergai O, Dergai M, Skrypkina I, et al. (2008) Alternative splicing affecting the SH3A domain controls the binding properties of intersectin 1 in neurons. Biochem Biophys Res Commun 372: 929–934.
[25]  Nikolaienko O, Skrypkina I, Tsyba L, Fedyshyn Y, Morderer D, et al. (2009) Intersectin 1 forms a complex with adaptor protein Ruk/CIN85 in vivo independently of epidermal growth factor stimulation. Cell Signal 21: 753–759.
[26]  Pucharcos C, Estvill X, de la Luna S (2000) Intersectin 2, a new multimodular protein involved in clathrin-mediated endocytosis. FEBS Lett 478: 43–51.
[27]  Aranda S, Alvarez M, Turro S, Laguna A, de la Luna S (2008) Sprouty2-mediated inhibition of fibroblast growth factor signaling is modulated by the protein kinase DYRK1A. Mol Cell Biol 28: 5899–5911.
[28]  Dergai O, Novokhatska O, Dergai M, Skrypkina I, Tsyba L, et al. (2010) Intersectin 1 forms complexes with SGIP1 and Reps1 in clathrin-coated pits. Biochem Biophys Res Commun 402: 408–413.
[29]  Colaluca IN, Tosoni D, Nuciforo P, Senic-Matuglia F, Galimberti V, et al. (2008) NUMB controls p53 tumour suppressor activity. Nat 451: 76–80.
[30]  Santonico E, Panni S, Falconi M, Castagnoli L, Cesareni G (2007) Binding to DPF-motif by the POB1 EH domain is responsible for POB1-Eps15 interaction. BMC Biochem 8: 29.
[31]  Dergai M, Skrypkina I, Dergai O, Tsyba L, Novokhatska O, et al. (2011) Identification and characterization of a novel mammalian isoform of the endocytic adaptor ITSN1. Gene 485: 120–129.
[32]  Obenauer JC, Cantley LC, Yaffe MB (2003) Scansite 2.0: Proteome-wide prediction of cell signaling interactions using short sequence motifs. Nucleic Acids Res 31: 3635–3641.
[33]  Sali A, Blundell TL (1993) Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol 234: 779–815.
[34]  Chen VB, Arendall WB, Headd JJ, Keedy DA, Immormino RM, et al. (2010) MolProbity: all-atom structure validation for macromolecular crystallography. Acta Cryst D66: 12–21.
[35]  Wong KA, Wilson J, Russo A, Wang L, Okur MN, et al. (2012) Intersectin (ITSN) family of scaffolds function as molecular hubs in protein interaction networks. PLoS One 7: e36023.
[36]  Morderer D, Nikolaienko O, Skrypkina I, Cherkas V, Tsyba L, et al. (2012) Endocytic adaptor protein intersectin 1 forms a complex with microtubule stabilizer STOP in neurons. Gene 505: 360–364.
[37]  Dergai O, Dergai M, Skrypkina I, Matskova L, Tsyba L, et al. (2013) The LMP2A protein of Epstein-Barr virus regulates phosphorylation of ITSN1 and Shb adaptors by tyrosine kinases. Cell Signal 25: 33–40.
[38]  Larson SF, Davidson AR (2000) The identification of conserved interactions within the SH3 domain by alignment of sequences and structures. Prot Sci 9: 2170–2180.
[39]  Dergai M, Tsyba L, Dergai O, Zlatskii I, Skrypkina I, et al. (2010) Microexon-based regulation of ITSN1 and Src SH3 domains specificity relies on introduction of charged amino acids into the interaction interface. Biochem Biophys Res Commun 399: 307–312.
[40]  Lim WA, Richards FM, Fox RO (1994) Structural determinants of peptide-binding orientation and of sequence specificity in SH3 domains. Nat 372: 375–379.
[41]  Feng S, Chen JK, Yu H, Simon JA, Schreiber SL (1994) Two binding orientations for peptides to the Src SH3 domain: development of a general model for SH3-ligand interactions. Sci 266: 1241–1247.
[42]  Betts MJ, Russell RB (2003) Amino acid properties and consequences of subsitutions. In: Barnes MR, Gray IC, editors. Bioinformatics for Geneticists. Wiley. 289–314.
[43]  Primeau M, Ben Djoudi Ouadda A, Lamarche-Vane N (2011) Cdc42 GTPase-activating protein (CdGAP) interacts with the SH3D domain of Intersectin through a novel basic-rich motif. FEBS Let 585: 847–853.
[44]  Hornbeck PV, Kornhauser JM, Tkachev S, Zhang B, Skrzypek E, et al. (2012) PhosphoSitePlus: a comprehensive resource for investigating the structure and function of experimentally determined post-translational modifications in man and mouse. Nucleic Acids Res 40: D261–270.
[45]  Wang LH, Rothberg KG, Anderson RG (1993) Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J Cell Biol 123: 1107–1117.
[46]  Ahmad KF, Lim WA (2010) The minimal autoinhibited unit of the guanine nucleotide exchange factor intersectin. PLoS ONE 5: e11291.
[47]  Harkiolaki M, Gilbert RJC, Jones EY, Feller SM (2006) The C-terminal SH3 domain of CRKL as a dynamic dimerization module transiently exposing a nuclear export signal. Structure 14: 1741–1753.
[48]  Nishida M, Nagata K, Hachimori Y, Horiuchi M, Ogura K, et al. (2001) Novel recognition mode between Vav and Grb2 SH3 domains. EMBO J 20: 2995–3007.
[49]  Huang PH, Mukasa A, Bonavia R, Flynn RA, Brewer ZE, et al. (2007) Quantitative analysis of EGFRvIII cellular signaling networks reveals a combinatorial therapeutic strategy for glioblastoma. Proc Natl Acad Sci USA 104: 12867–12872.
[50]  Neduva V, Russell RB (2005) Linear motifs: Evolutionary interaction switches. FEBS Lett 579: 3342–3345.

Full-Text

comments powered by Disqus