Parvin is a putative F-actin binding protein important for integrin-mediated cell adhesion. Here we used overexpression of Drosophila Parvin to uncover its functions in different tissues in vivo. Parvin overexpression caused major defects reminiscent of metastatic cancer cells in developing epithelia, including apoptosis, alterations in cell shape, basal extrusion and invasion. These defects were closely correlated with abnormalities in the organization of F-actin at the basal epithelial surface and of integrin-matrix adhesion sites. In wing epithelium, overexpressed Parvin triggered increased Rho1 protein levels, predominantly at the basal side, whereas in the developing eye it caused a rough eye phenotype and severely disrupted F-actin filaments at the retina floor of pigment cells. We identified genes that suppressed these Parvin-induced dominant effects, depending on the cell type. Co-expression of both ILK and the apoptosis inhibitor DIAP1 blocked Parvin-induced lethality and apoptosis and partially ameliorated cell delamination in epithelia, but did not rescue the elevated Rho1 levels, the abnormal organization of F-actin in the wing and the assembly of integrin-matrix adhesion sites. The rough eye phenotype was suppressed by coexpression of either PTEN or Wech, or by knock-down of Xrp1. Two main conclusions can be drawn from our studies: (1), high levels of cytoplasmic Parvin are toxic in epithelial cells; (2) Parvin in a dose dependent manner affects the organization of actin cytoskeleton in both wing and eye epithelia, independently of its role as a structural component of the ILK-PINCH-Parvin complex that mediates the integrin-actin link. Thus, distinct genetic interactions of Parvin occur in different cell types and second site modifier screens are required to uncover such genetic circuits.
References
[1]
Bokel C, Brown N (2002) Integrins in development: moving on, responding to, and sticking to the extracellular matrix. Dev Cell 3: 311–321.
[2]
Geiger B, Yamada KM (2011) Molecular Architecture and Function of Matrix Adhesions Cold Spring Harb Perspect Biol. 3: a005033.
[3]
Sepulveda JL, Wu C (2006) The parvins. Cell Mol Life Sci 63: 25–35.
[4]
Wickstrom SA, Lange A, Montanez E, Fassler R (2010) The ILK/PINCH/parvin complex: the kinase is dead, long lived the pseudokinase! EMBO J. 29: 281–291.
[5]
Montanez E, Wickstr？m SA, Altst？tter J, Chu H, F？ssler R (2009) Alpha-parvin controls vascular mural cell recruitment to vessel wall by regulating RhoA/ROCK signalling. EMBO J 28: 3132–3144.
[6]
Vakaloglou KM, Chountala M, Zervas CG (2012) Functional analysis of parvin and different modes of IPP-complex assembly at integrin sites during Drosophila development. J Cell Sci 125: 3221–3232.
[7]
Johnstone CN, Mongroo PS, Rich AS, Schupp M, Bowser MJ, et al. (2008) Parvin-beta inhibits breast cancer tumorigenicity and promotes CDK9-mediated peroxisome proliferator-activated receptor gamma 1 phosphorylation. Mol Cell Biol 28: 687–704.
[8]
Zervas CG, Gregory SL, Brown NH (2001) Drosophila integrin-linked kinase is required at sites of integrin adhesion to link the cytoskeleton to the plasma membrane. J Cell Biol 152: 1007–1018.
[9]
Gutzeit HO, Eberhardt W, Gratwohl E (1991) Laminin and basement membrane-associated microfilaments in wild-type and mutant Drosophila ovarian follicles. J Cell Sci 100: 781–788.
[10]
Fukuda T, Guo L, Shi X, Wu C (2003) CH-ILKBP regulates cell survival by facilitating the membrane translocation of protein kinase B/Akt. J Cell Biol 160: 1001–1008.
[11]
Zhang Y, Chen X, Tu Y, Wu C (2004) Distinct roles of two structurally closely related focal adhesion proteins, a-parvins and b-parvins, in regulation of cell morphology and survival. J Biol Chem 279: 41695–41705.
[12]
Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118: 401–415.
[13]
Fan YaB, A. (2010 ) The cleaved-Caspase-3 antibody is a marker of Caspase-9-like DRONC activity in Drosophila. Cell Death Differ 17: 534–539.
[14]
Adachi-Yamada T, Fujimura-Kamada K, Nishida Y, Matsumoto K (1999) Distortion of proximodistal information causes JNK-dependent apoptosis in Drosophila wing. Nature 400: 166–169.
[15]
Agnès F, Suzanne M, Noselli S (1999) The Drosophila JNK pathway controls the morphogenesis of imaginal discs during metamorphosis. Development 126: 5453–5462.
[16]
Wang SL, Hawkins CJ, Yoo SJ, Müller HA, Hay BA (1999) The Drosophila caspase inhibitor DIAP1 is essential for cell survival and is negatively regulated by HID. Cell 98: 453–463.
[17]
Page-McCaw A, Serano J, Sante JM, Rubin GM (2003) Drosophila matrix metalloproteinases are required for tissue remodeling, but not embryonic development. Dev Cell 4: 95–106.
[18]
Brown NH, Gregory SL, Rickoll WL, Fessler LI, Prout M, et al. (2002) Talin is essential for integrin function in Drosophila. Dev Cell 3: 569–579.
[19]
Domínguez-Giménez P, Brown N, Martín-Bermudo M (2007) Integrin-ECM interactions regulate the changes in cell shape driving the morphogenesis of the Drosophila wing epithelium. J Cell Sci 120: 1061–1071.
[20]
Breitsprecher D, Kiesewetter AK, Linkner J, Vinzenz M, Stradal TE, et al. (2011) Molecular mechanism of Ena/VASP-mediated actin-filament elongation. EMBO J 30: 456–467.
[21]
Becam I, Rafel N, Hong X, Cohen SM, Milán M (2011) Notch-mediated repression of bantam miRNA contributes to boundary formation in the Drosophila wing. Development 138: 3781–3789.
[22]
Thiery JP, Acloque H, Huang RY, Nieto MA (2009) Epithelial-mesenchymal transitions in development and disease. Cell 139: 871–890.
[23]
Berganti？os C, Corominas M, Serras F (2010) Cell death-induced regeneration in wing imaginal discs requires JNK signalling. Development 137: 1169–1179.
[24]
Li W, Baker N (2007) Engulfment is required for cell competition. Cell 129: 1215–1225.
[25]
Neisch AL, Speck O, Stronach B, Fehon RG (2010) Rho1 regulates apoptosis via activation of the JNK signaling pathway at the plasma membrane. J Cell Biol 189: 311–323.
[26]
Vidal M, Larson DE, Cagan RL (2006) Csk-deficient boundary cells are eliminated from normal Drosophila epithelia by exclusion, migration, and apoptosis. Dev Cell 10: 33–44.
[27]
Grzeschik NA, Knust E (2005) IrreC/rst-mediated cell sorting during Drosophila pupal eye development depends on proper localisation of DE-cadherin Development. 132: 2035–2045.
[28]
Lin DMaG, C S (1994) Ectopic and increased expression of Fasciclin II alters motoneuron growth cone guidance. Neuron 13: 507–523.
[29]
Tomlinson A, Bowtell DD, Hafen E, Rubin GM (1987) Localization of the sevenless protein, a putative receptor for positional information, in the eye imaginal disc of Drosophila. Cell 51: 143–150.
[30]
Cagan R (2009) Principles of Drosophila eye differentiation. Curr Top Dev Biol 89: 115–135.
[31]
Longley RL, Ready DF (1995) Integrins and the development of three-dimensional structure in the Drosophila compound eye. Dev Biol 171: 415–433.
[32]
Martin-Bermudo MD, Dunin-Borkowski OM, Brown NH (1997) Specificity of PS integrin function during embryogenesis resides in the a subunit extracellular domain. EMBO 16: 4184–4193.
[33]
L？er B, Bauer R, Bornheim R, Grell J, Kremmer E, et al. (2008) The NHL-domain protein Wech is crucial for the integrin-cytoskeleton link. Nat Cell Biol 10: 422–428.
[34]
Staveley BE, Ruel L, Jin J, Stambolic V, Mastronardi FG, et al. (1998) Genetic analysis of protein kinase B (AKT) in Drosophila. Current Biology 8: 599–602.
[35]
Huang H, Potter CJ, Tao W, Li DM, Brogiolo W, et al. (1999) PTEN affects cell size, cell proliferation and apoptosis during Drosophila eye development. Development 126: 5365–5372.
[36]
Dietzl G, Chen D, Schnorrer F, Su KC, Barinova Y, et al. (2007) A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448: 151–156.
[37]
Lee SB, Cho KS, Kim E, Chung J (2003) blistery encodes Drosophila tensin protein and interacts with integrin and the JNK signaling pathway during wing development. Development 130: 4001–4010.
[38]
Chen GC, Turano B, Ruest PJ, Hagel M, Settleman J, et al. (2005) Regulation of Rho and Rac signaling to the actin cytoskeleton by paxillin during Drosophila development. Mol Cell Biol 25: 979–987.
[39]
Mishima W, Suzuki A, Yamaji S, Yoshimi R, Ueda A, et al. (2004) The first CH domain of affixin activates Cdc42 and Rac1 through alphaPIX, a Cdc42/Rac1-specific guanine nucleotide exchanging factor. Genes Cells 9: 193–204.
[40]
LaLonde DP, Grubinger M, Lamarche-Vane N, Turner CE (2006) CdGAP Associates with Actopaxin to Regulate Integrin-Dependent Changes in Cell Morphology and Motility. Curr Biol 16: 1375–1385.
[41]
Widmann TJ, Dahmann C (2009) Dpp signaling promotes the cuboidal-to-columnar shape transition of Drosophila wing disc epithelia by regulating Rho1. J Cell Sci 122: 1362–1373.
[42]
Speck O, Hughes SC, Noren NK, Kulikauskas RM, Fehon RG (2003) Moesin functions antagonistically to the Rho pathway to maintain epithelial integrity. Nature 421: 83–87.
[43]
Fan Y, Bergmann A (2008) Apoptosis-induced compensatory proliferation. The Cell is dead. Long live the Cell! Trends in Cell Biol 18: 467–473.
[44]
Janody F, Treisman JE (2006) Actin capping protein alpha maintains vestigial-expressing cells within the Drosophila wing disc epithelium. Development 133: 3349–3357.
[45]
Scanga SE, Ruel L, Binari RC, Snow B, Stambolic V, et al. (2000) The conserved PI3′K/PTEN/Akt signaling pathway regulates both cell size and survival in Drosophila. Oncogene 19: 3971–3977.
[46]
Kramer JM, Staveley BE (2003) GAL4 causes developmental defects and apoptosis when expressed in the developing eye of Drosophila melanogaster. Genet Mol Res 2: 43–47.
[47]
Ellis MC, O’Neill EM, Rubin GM (1993) Expression of Drosophila glass protein and evidence for negative regulation of its activity in non-neuronal cells by another DNA-binding protein. Development 119: 855–865.
[48]
Kimura M, Murakami T, Kizaka-Kondoh S, Itoh M, Yamamoto K, et al. (2010) Functional molecular imaging of ILK-mediated Akt/PKB signaling cascades and the associated role of beta-parvin. J Cell Sci 123: 747–755.
[49]
Radimerski T, Montagne J, Hemmings-Mieszczak M, Thomas G (2002) Lethality of Drosophila lacking TSC tumor suppressor function rescued by reducing dS6K signaling. Genes Dev 16: 2627–2632.
[50]
Akdemir F, Christich A, Sogame N, Chapo J, Abrams JM (2007) p53 directs focused genomic responses in Drosophila. Oncogene 26: 5184–5193.
[51]
Jani K, Sch？ck F (2007) Zasp is required for the assembly of functional integrin adhesion sites. J Cell Biol 179: 1583–1597.
