Summary
A series of transposon-induced optic morphology (Om) mutants found in a hypermutable marker stock of Drosophila ananassae provides a useful system for analyzing the molecular mechanism of eye morphogenesis. In the present study, one of the 25 Om loci so far reported, Om(2D), has been subjected to histological and molecular analyses as a first step toward understanding the role of Om genes in eye morphogenesis. Histological abnormalities observed during eye morphogenesis of the mutant, i.e. cell death within the eye-antennal discs of third instar larvae, and loss of the lamina, disorganized ommatidia and atrophied optic lobes in adults, were all comparable to those reported with various eye morphology mutants of D. melanogaster. Approximately 25 kb of genomic DNA including the Om(2D) locus was cloned by tom tagging. Southern blot and cloning analyses of two alleles of the Om(2D) locus revealed that insertions of the tom element occurred at three sites within 359 bp; two tandemly arrayed toms sharing one long terminal repeat at the junction and an internally deleted tom were present 359 by apart from each other in Om(2D)63, while a single tom in reverse orientation was present within the 359 by in Om (2D)10a. Host DNA sequences at the three insertion sites were TATAT or AATAT, and ATAT was duplicated upon the tom insertion. Three spontaneous revertants and one induced extreme derivative of Om(2D)63 were obtained and characterized. A complete revertant lost all the preexisting tom elements. Two partial revertants lost one or two of the preexisting tom elements. In the extreme derivative, an additional insertion sequence was found within the two tandem tom elements. Northern blot analysis showed two transcription units in the Om (2D) region: one was on the centromere side of the tom insertion site and expressed a 3.2 kb major RNA and several minor RNAs; the other resided on the telomere side of the tom insertion and expressed a 1.5 kb RNA. Both 3.2 kb and 1.5 kb transcripts were expressed throughout development, but the former was more abundant in mutant embryos and the latter more highly expressed in mutant third instar larvae than in the corresponding stages in wild type.
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Bingham PM, Levis R, Rubin GM (1981) Cloning of DNA sequences from the white locus of Drosophila melanogaster by a novel and general method. Cell 25:693–704
Campos-Ortega JA (1980) On compound eye development in Drosophila melanogaster. Curr Top Dev Biol 15:347–371
Campuzano S, Balcells L, Villares R, Carramolino L, Garcia-Alonso L, Modolell J (1986) Excess function Hairy-wing mutations caused by gypsy and copia insertions within structural genes of the achaete-scute locus of Drosophila. Cell 44:303–312
Davis PS, Shen MW, Judd BH (1987) Asymmetrical pairings of transposons in and proximal to the white locus of Drosophila account for four classes of regularly occurring exchange products. Proc Natl Acad Sci USA 84:174–178
Engels WR, Preston CR, Thompson P, Eggleston WB (1986) In situ hybridization to Drosophila salivary chromosomes with biotinylated DNA probes and alkaline phosphatase. Focus 8(1):6–8
Fischbach KF (1983) Neural cell types surviving congenital sensory deprivation in the optic lobes of Drosophila melanogaster. Dev Biol 195:1–18
Frischauf A-M, Lehrach H, Poustka A, Murray N (1983) Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170:827–842
Fristrom D (1969) Cellular degeneration in the production of some mutant phenotypes in Drosophila melanogaster. Mol Gen Genet 103:363–379
Geyer PK, Spana C, Corces V (1986) On the molecular mechanism of gypsy-induced mutations at the yellow locus of Drosophila melanogaster. EMBO J 5:2657–2662
Goldberg ML, Sheen J-Y, Gehring WJ, Green MM (1983) Unequal crossing-over associated with asymmetrical synapsis between nomadic elements in the Drosophila melanogaster Genome. Proc Natl Acad Sci USA 80:5017–5021
Hartman H, Hayes TL (1971) Scanning electron microscopy of Drosophila. J Hered 62:41–44
Hattori M, Sakaki Y (1986) Dideoxy sequencing method using denatured plasmid templates. Anal Biochem 152:232–238
Henkemeyer MJ, Gertler FB, Goodman W, Hoffman FM (1987) The Drosophila abelson proto-oncogene homolog; identification of mutant alleles that have pleiotropic effects late in development. Cell 51:821–828
Hinton CW (1980) New mutants in Drosophila ananassae. Drosophila Inf Serv 55:213–216
Hinton CW (1984) Morphogenetically specific mutability in Drosophila ananassae. Genetics 106:631–653
Hinton CW (1988) Formal relations between Om mutants and their suppressors in Drosophila ananassae. Genetics 120:1035–1042
Hofbauer A, Campos-Ortega JA (1976) Cell clones and pattern formation: Genetic eye mosaics in Drosophila melanogaster. Roux's Arch Dev Biol 178:275–289
Hong GF (1982) A systematic DNA sequencing strategy. J Mol Biol 158:539–549
Hori SH, Akasaka M, Ito H, Hanaoka T, Tanda S, Ohtsuka E, Miura K, Takahashi T, Tang JJN (1985) Cloning of the glucose-6-phosphate dehydrogenase gene of Drosophila melanogaster using 17-base oligonucleotide mixtures as probes. Jpn J Genet 60:455–463
Kushida H, Kushida T, Iijima H (1985) An improved method for both light and electron microscopy of identical sites in semi-thin sections under 200 kV transmission electron microscope. J Electron Microsc 34:438–441
Lindsley DL, Grell EH (1968) Genetic variations in Drosophila melanogaster. Carnegie Inst Wash Publ 627
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Matsubayashi H, Matsuda M, Tobari YN (1986) The transposable element tom, insertion mutation Om and suppressors of Om in Drosophila ananassae (abstract, in Japanese). Jpn J Genet 61:594
Meyerowitz EM, Kankel DR (1978) A genetic analysis of visual system development in Drosophila melanogaster. Dev Biol 62:112–142
Mizrokhi LJ, Obolenkova LA, Priimagi AF, Ilyin YV, Gerasimova TI, Georgiev GP (1985) The nature of unstable insertion mutations and reversions in the locus cut of Drosophila melanogaster: molecular mechanism of transposition memory. EMBO J 4:3781–3787
Moriwaki D, Ito S (1969) Studies on puffing in the salivary gland chromosomes of Drosophila ananassae. Jpn J Genet 44:129–138
Pilkington RW (1941) Facet mutants of Drosophila. Proc Zool Soc London A 111:199–222
Pirrotta V (1986) Cloning Drosophila genes. In: Roberts DB (ed) Drosophila: A practical approach. IRL Press, Oxford, pp 83–110
Power ME (1943) The effect of reduction in numbers of ommatidia upon the brain of Drosophila melanogaster. J Exp Zool 94:33–71
Ranson R (1979) The time of action of the three mutations affecting Drosophila eye morphogenesis. J Embryo Exp Morph 53:225–235
Saint R, Kalionis B, Lockett TJ, Elizurl A (1988) Pattern formation in the developing eye of Drosophila melanogaster is regulated by the homooo-box gene, rough. Nature 334:151–154
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Shrimpton AE, Montgomery EA, Langley CH (1986) Om mutations in Drosophila ananassae are linked to insertions of a transposable element. Genetics 114:125–135
Swaroop A, Paco-Larson ML, Garen A (1985) Molecular genetics of a transposon-induced dominant mutation in the Drosophila locus Glued. Proc Natl Acad Sci USA 82:1751–1755
Tanda S, Shrimpton AE, Ling-Ling C, Itayama H, Matsubayashi H, Saigo K, Tobari YN, Langley CH (1988) Retrovirus-like features and site specific insertions of a transposable element, tom, in Drosophila ananassae. Mol Gen Genet 214:405–411
Tanda S, Shrimpton AE, Hinton CW, Langley CH (1989) Analysis of the Om(ID) locus in Drosophila ananassae. Genetics 123:495–502
Tomlinson A, Kimmel BE, Rubin GM (1988) rough, a Drosophila homeobox gene required in photoreceptors R2 and R5 for inductive interactions in the developing eye. Cell 55:771–784
Varmus HE (1984) The molecular genetics of cellular oncogenes. Annu Rev Genet 18553–612
Zachar Z, Davison D, Garza D, Bingham PM (1985) A detailed developmental and structural study of the transcriptional effects of insertion of the copia transposon into the white locus of Drosophila melanogaster. Genetics 111:495–515
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Communicated by B.H. Judd
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Matsubayashi, H., Juni, N., Usui, K. et al. Molecular and histological characterizations of the Om(2D) mutants in Drosophila ananassae . Molec. Gen. Genet. 227, 165–172 (1991). https://doi.org/10.1007/BF00259667
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DOI: https://doi.org/10.1007/BF00259667