Characterization of Drosophila mini-me, a Gene Required for Cell Proliferation and Survival

In the developing Drosophila eye, the morphogenetic furrow is a developmental organizing center for patterning and cell proliferation. The furrow acts both to limit eye size and to coordinate the number of cells to the number of facets. Here we report the molecular and functional characterization of Drosophila mini-me (mnm), a potential regulator of cell proliferation and survival in the developing eye. We first identified mnm as a dominant modifier of hedgehog loss-of-function in the developing eye. We report that mnm encodes a conserved protein with zinc knuckle and RING finger domains. We show that mnm is dispensable for patterning of the eye disc, but required in the eye for normal cell proliferation and survival. We also show that mnm null mutant cells exhibit altered cell cycle profiles and contain excess nucleic acid. Moreover, mnm overexpression can induce cells to proliferate and incorporate BrdU. Thus, our data implicate mnm as a regulator of mitotic progression during the proliferative phase of eye development, possibly through the control of nucleic acid metabolism.

[1]  B. Dickson,et al.  Analysis of Drosophila photoreceptor axon guidance in eye-specific mosaics. , 2000, Development.

[2]  Jean-Marie Schmitter,et al.  Mpe1, a Zinc Knuckle Protein, Is an Essential Component of Yeast Cleavage and Polyadenylation Factor Required for the Cleavage and Polyadenylation of mRNA , 2001, Molecular and Cellular Biology.

[3]  U. Heberlein,et al.  Mechanisms of drosophila retinal morphogenesis: The virtues of being progressive , 1995, Cell.

[4]  R. Scott,et al.  The proliferation potential protein-related (P2P-R) gene with domains encoding heterogeneous nuclear ribonucleoprotein association and Rb1 binding shows repressed expression during terminal differentiation. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[5]  G. Rubin,et al.  The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes. , 1999, Genetics.

[6]  T. Lecuit,et al.  Mad acts downstream of Dpp receptors, revealing a differential requirement for dpp signaling in initiation and propagation of morphogenesis in the Drosophila eye. , 1996, Development.

[7]  R. Cagan,et al.  The emergence of order in the Drosophila pupal retina. , 1989, Developmental biology.

[8]  J. Mohler Requirements for hedgehog, a segmental polarity gene, in patterning larval and adult cuticle of Drosophila. , 1988, Genetics.

[9]  V. Rotter,et al.  PACT: cloning and characterization of a cellular p53 binding protein that interacts with Rb , 1997, Oncogene.

[10]  I. Hariharan,et al.  Uncoupling Cell Fate Determination from Patterned Cell Division in the Drosophila Eye , 1995, Science.

[11]  S. Higashijima,et al.  Dual Bar homeo box genes of Drosophila required in two photoreceptor cells, R1 and R6, and primary pigment cells for normal eye development. , 1992, Genes & development.

[12]  N. Baker,et al.  Extracellular signals responsible for spatially regulated proliferation in the differentiating Drosophila eye. , 2005, Developmental cell.

[13]  P. Ingham,et al.  hedgehog is required for the proliferation and specification of ovarian somatic cells prior to egg chamber formation in Drosophila. , 1996, Development.

[14]  S. Andersson,et al.  RK2, a glial-specific homeodomain protein required for embryonic nerve cord condensation and viability in Drosophila. , 1994, Development.

[15]  P. Beachy,et al.  Secretion and localized transcription suggest a role in positional signaling for products of the segmentation gene hedgehog , 1992, Cell.

[16]  T Wolff,et al.  Cell death in normal and rough eye mutants of Drosophila. , 1991, Development.

[17]  G M Rubin,et al.  wingless inhibits morphogenetic furrow movement in the Drosophila eye disc. , 1995, Development.

[18]  T Marty,et al.  Ectopic cyclin E expression induces premature entry into S phase and disrupts pattern formation in the Drosophila eye imaginal disc. , 1995, Development.

[19]  C. Nüsslein-Volhard,et al.  Mutations affecting segment number and polarity in Drosophila , 1980, Nature.

[20]  N. Perrimon,et al.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. , 1993, Development.

[21]  Y Taya,et al.  cDNA sequence and chromosomal localization of a novel human protein, RBQ-1 (RBBP6), that binds to the retinoblastoma gene product. , 1995, Genomics.

[22]  U. Banerjee,et al.  The little R cell that could. , 2004, International Journal of Developmental Biology.

[23]  Y. H. Sun,et al.  Localized Notch signal acts through eyg and upd to promote global growth in Drosophila eye , 2004, Development.

[24]  P. Freemont Ubiquitination: RING for destruction? , 2000, Current Biology.

[25]  C. Heldin,et al.  The L45 loop in type I receptors for TGF‐β family members is a critical determinant in specifying Smad isoform activation , 1998, FEBS letters.

[26]  Antonio Baonza,et al.  Control of cell proliferation in the Drosophila eye by Notch signaling. , 2005, Developmental cell.

[27]  K. Roth,et al.  In situ immunodetection of activated caspase-3 in apoptotic neurons in the developing nervous system , 1998, Cell Death and Differentiation.

[28]  G. Rubin,et al.  Analysis of the cis-acting requirements for germ-line-specific splicing of the P-element ORF2-ORF3 intron. , 1989, Genes & development.

[29]  N. Baker,et al.  The EGF Receptor Defines Domains of Cell Cycle Progression and Survival to Regulate Cell Number in the Developing Drosophila Eye , 2001, Cell.

[30]  G. Rubin,et al.  Analysis of genetic mosaics in developing and adult Drosophila tissues. , 1993, Development.

[31]  S. Zipursky,et al.  Cell cycle progression in the developing Drosophila eye: roughex encodes a novel protein required for the establishment of G1 , 1994, Cell.

[32]  S. Selleck,et al.  Regulation of Cell Cycle Synchronization by decapentaplegic During Drosophila Eye Development , 1997, Science.

[33]  R. Scott,et al.  P2P-R deficiency modifies nocodazole-induced mitotic arrest and UV-induced apoptosis. , 2002, Anticancer research.

[34]  M. Freeman,et al.  Cell determination strategies in the Drosophila eye. , 1997, Development.

[35]  Y. Jan,et al.  Genghis Khan (Gek) as a putative effector for Drosophila Cdc42 and regulator of actin polymerization. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[36]  P. Ingham,et al.  Hedgehogs tryst with the cell cycle , 2002, Journal of Cell Science.

[37]  K. Moses,et al.  The segment polarity gene hedgehog is required for progression of the morphogenetic furrow in the developing Drosophila eye , 1993, Cell.

[38]  N. Baker Cell proliferation, survival, and death in the Drosophila eye. , 2001, Seminars in cell & developmental biology.

[39]  I. Hariharan,et al.  An overexpression screen in Drosophila for genes that restrict growth or cell-cycle progression in the developing eye. , 2002, Genetics.

[40]  G. Struhl,et al.  Progression of the morphogenetic furrow in the Drosophila eye: the roles of Hedgehog, Decapentaplegic and the Raf pathway. , 1999, Development.

[41]  W. Fu,et al.  Deciphering synergistic and redundant roles of Hedgehog, Decapentaplegic and Delta that drive the wave of differentiation in Drosophila eye development , 2003, Development.

[42]  Wei Du,et al.  Hedgehog regulates cell growth and proliferation by inducing Cyclin D and Cyclin E , 2002, Nature.

[43]  Lawrence Lum,et al.  The Hedgehog Response Network: Sensors, Switches, and Routers , 2004, Science.

[44]  I. Hariharan,et al.  A Cyclin-Dependent Kinase Inhibitor, Dacapo, Is Necessary for Timely Exit from the Cell Cycle during Drosophila Embryogenesis , 1996, Cell.

[45]  A. Tomlinson,et al.  Cellular interactions in the developing Drosophila eye. , 1988, Development.

[46]  A. Preiss,et al.  Notchspl is deficient for inductive processes in the eye, and E(spl)D enhances split by interfering with proneural activity. , 1999, Developmental biology.

[47]  G. Mardon,et al.  R8 development in the Drosophila eye: a paradigm for neural selection and differentiation. , 2002, Development.

[48]  Gerald M. Rubin,et al.  The glass gene encodes a zinc-finger protein required by Drosophila photoreceptor cells , 1989, Nature.

[49]  Gerald M. Rubin,et al.  The activities of two Ets-related transcription factors required for drosophila eye development are modulated by the Ras/MAPK pathway , 1994, Cell.

[50]  Tian Xu,et al.  A Genetic Screen in Drosophila for Metastatic Behavior , 2003, Science.

[51]  Y. Jan,et al.  Patterns of expression of cut, a protein required for external sensory organ development in wild-type and cut mutant Drosophila embryos. , 1990, Genes & development.

[52]  C. K. Motzny,et al.  The Drosophila cubitus interruptus protein and its role in the wingless and hedgehog signal transduction pathways , 1995, Mechanisms of Development.

[53]  B. Dickson,et al.  The Drosophila Tuberous Sclerosis Complex Gene Homologs Restrict Cell Growth and Cell Proliferation , 2001, Cell.

[54]  K. Arndt,et al.  Coiled Coil Domains: Stability, Specificity, and Biological Implications , 2004, Chembiochem : a European journal of chemical biology.

[55]  Christian Dahmann,et al.  Extrusion of Cells with Inappropriate Dpp Signaling from Drosophila Wing Disc Epithelia , 2005, Science.

[56]  J P Kumar,et al.  Dissecting the roles of the Drosophila EGF receptor in eye development and MAP kinase activation. , 1998, Development.

[57]  N. Perrimon,et al.  Extrusion and Death of DPP/BMP-Compromised Epithelial Cells in the Developing Drosophila Wing , 2005, Science.

[58]  S. Zipursky,et al.  Induction of Drosophila eye development by decapentaplegic. , 1997, Development.

[59]  I. Rebay,et al.  Signal integration during development: Insights from the Drosophila eye , 2004, Developmental dynamics : an official publication of the American Association of Anatomists.

[60]  G. Mardon,et al.  senseless Repression of rough Is Required for R8 Photoreceptor Differentiation in the Developing Drosophila Eye , 2001, Neuron.

[61]  A. Tomlinson The cellular dynamics of pattern formation in the eye of Drosophila. , 1985, Journal of embryology and experimental morphology.

[62]  Yves Grau,et al.  atonal is a proneural gene that directs chordotonal organ formation in the Drosophila peripheral nervous system , 1993, Cell.

[63]  J. Horsfield,et al.  decapentaplegic is required for arrest in G1 phase during Drosophila eye development. , 1998, Development.

[64]  G. Rubin,et al.  Genetic transformation of Drosophila with transposable element vectors. , 1982, Science.

[65]  S. Zipursky,et al.  The bride of sevenless and sevenless interaction: Internalization of a transmembrane ligand , 1992, Cell.

[66]  I. Hariharan,et al.  Expression of the cyclin-dependent kinase inhibitor Dacapo is regulated by Cyclin E , 2000, Mechanisms of Development.

[67]  S. Benzer,et al.  Development of the Drosophila retina, a neurocrystalline lattice. , 1976, Developmental biology.

[68]  M. Mlodzik,et al.  Morphogenetic furrow initiation and progression during eye development in Drosophila: the roles of decapentaplegic, hedgehog and eyes absent. , 2000, Development.

[69]  G M Rubin,et al.  Expression of baculovirus P35 prevents cell death in Drosophila. , 1994, Development.

[70]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[71]  Marek Mlodzik,et al.  The regulation of hedgehog and decapentaplegic during Drosophila eye imaginal disc development , 1996, Mechanisms of Development.

[72]  C. Lehner,et al.  Synergistic action of Drosophila cyclins A and B during the G2‐M transition. , 1993, The EMBO journal.

[73]  M. Summers Zinc finger motif for single‐stranded nucleic acids? investigations by nuclear magnetic resonance , 1991, Journal of cellular biochemistry.

[74]  Stephen C. Ekker,et al.  The product of hedgehog autoproteolytic cleavage active in local and long-range signalling , 1995, Nature.

[75]  R. Scott,et al.  P2P‐R protein localizes to the nucleolus of interphase cells and the periphery of chromosomes in mitotic cells which show maximum P2P‐R immunoreactivity , 2002, Journal of cellular physiology.

[76]  M. Dominguez,et al.  Growth and specification of the eye are controlled independently by Eyegone and Eyeless in Drosophila melanogaster , 2004, Nature Genetics.

[77]  The Drosophila TGF alpha homolog Spitz acts in photoreceptor recruitment in the developing retina. , 1997, Development.

[78]  K. Moses,et al.  The Drosophila TGF alpha homolog Spitz acts in photoreceptor recruitment in the developing retina. , 1997, Development.