Abstract
The optomotor-blind gene of Drosophila melanogaster is large and genetically complex. Five partly independent complementation groups are uncovered by several viable and lethal mutations at the locus. At least 15 RNA signals have been detected by Northern blot analysis. One of them, T3, derived from a 75 kb primary transcript, has been proposed as the carrier of optomotor-blind function, based on the large size of its precursor and its tissue distribution. We here provide direct evidence that T3 is the optomotor-blind transcript. A facile and generally applicable selection scheme for the isolation of intragenic meiotic recombinants was applied to map two lethal optomotor-blind point mutations to exons of the T3 transcript. Amplification of mutant DNA by the polymerase chain reaction (PCR) and sequencing of the amplified exons revealed the presence of mutations that lead to truncation of the T3 open reading frame. The recombination rate observed in the optomotor-blind locus is within the range of rates that have been determined in a few other Drosophila loci.
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References
Ashburner M (1989) Drosophila. A laboratory handbook. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Banga SS, Bloomquist BT, Brodberg RK, Pye QN, Larrivee DC, Mason JM, Boyd JB, Pak WL (1986) Cytogenetic characterization of the 4BC region on the X chromosome of Drosophila melanogaster: Localization of the mei-9, norpA and omb genes. Chromosoma 93:341–346
Bausenwein B, Wolf R, Heisenberg M (1986) Genetic dissection of optomotor behavior in Drosophila melanogaster. Studies on wild-type and the mutant optomotor-blind H31 J Neurogenet 3:87–109
Begun DJ, Aquadro CF (1992) Levels of naturally occurring DNA polymorphism correlate with recombination rates in D. melanogaster. Nature 356:519–520
Blondeau J, Heisenberg M (1982) The three-dimensional optomotor torque system of Drosophila melanogaster. Studies on wildtype and the mutant optomotor-blind H31. J Comp Physiol 145:321–329
Brunner A, Wolf R, Pflugfelder GO, Poeck B, Heisenberg M (1992) Mutations in the proximal region of the optomotor-blind locus of Drosophila melanogaster reveal a gradient of neuroanatomical and behavioral phenotypes. J Neurogenet 8:43–55
Cribbs DL, Pultz MA, Johnson D, Mazzulla M, Kaufman TC (1992) Structural complexity and evolutionary conservation of the Drosophila homeotic gene proboscipedia. EMBO J 11:1437–1449
Doerig RE, Suter B, Gray M, Kubli E (1988) Identification of an amber nonsense mutation in the rosy516 gene by germline transformation of an amber suppressor tRNA gene. EMBO J 7:2579–2584
Dynan WS (1989) Modularity in promoters and enhancers. Cell 58:1–4
Fitzpatrick VD, Percival-Smith A, Ingles CJ, Krause HM (1992) Homeodomain-independent activity of the fushi tarazu polypeptide in Drosophila embryos. Nature 356:610–612
Gray M, Charpentier A, Walsh K, Wu P, Bender W (1991) Mapping point mutations in the Drosophila rosy locus using denaturing gradient gel blots. Genetics 127:139–149
Heisenberg M, Wonneberger R, Wolf R (1978) optomotor-blind H31. — a Drosophila mutant of the lobula plate giant neurons. J Comp Physiol 124:287–296
Hilliker AJ, Clark SH, Chovnick A (1991) The effect of DNA sequence of polymorphisms on intragenic recombination in the rosy locus of Drosophila melanogaster. Genetics 129:779–781
Hultman T, Stahl S, Hornes E, Uhlen M (1989) Direct solid-phase sequencing of genomic and plasmid DNA using magnetic beads as solid support. Nucleic Acids Res 17:4937–4946
Isoda K, Roth S, Nüsslein-Volhard C (1992) The functional domains of the Drosophila morphogen dorsal: evidence from the analysis of mutants. Genes Dev 6:619–630
Keohavong P, Thilly WG (1989) Fidelity of DNA polymerases in DNA amplification. Proc Natl Acad Sci USA 86:9253–9257
Kidd S, Lockett TJ, Young MW (1983) The Notch locus of Drosophila melanogaster. Cell 34:421–433
Lindsley DL, Grell EH (1968) Genetic variation in Drosophila melanogaster. Carnegie Inst Wash Publ 627
Lindsley DL, Zimm G (1987) The genome of Drosophila melanogaster. Part 3: Rearrangements. Drosophila Inf Serv 65
Liu S, McLeod E, Jack J (1991) Four distinct regulatory regions of the cut locus and their effect on cell type specification in Drosophila. Genetics 127:151–159
Morgan TH, Bridges CB (1916) Sex-linked inheritance in Drosophila. Carnegie Inst Wash Publ No 237
Pastink A, Heemskerk E, Nivard MJM, van Vliet CJ, Vogel EW (1991) Mutational specificity of ethyl methansulfonate in excision-repair-proficient and -deficient strains of Drosophila melanogaster. Mol Gen Genet 229:213–218
Peifer M, Wieschaus E (1990) The segment polarity gene armadillo encodes a functionally modular protein that is the Drosophila homolog of human plakoglobin. Cell 63:1167–1178
Pflugfelder GO, Schwarz H, Roth, Poeck B, Sigl A, Kerscher S, Jonschker B, Pak WL, Heisenberg M (1990) Genetic and molecular characterization of the optomotor-blind gene locus in Drosophila melanogaster. Genetics 126:91–104
Pflugfelder GO, Roth H, Poeck B, Kerscher S, Schwarz H, Jonschker B, Heisenberg M (1992a) The lethal(1)optomotor-blind gene of Drosophila melanogaster is a major organizer of optic lobe development: isolation and characterization of the gene. Proc Natl Acad Sci USA 89:1199–1203
Pflugfelder GO, Roth H, Poeck B (1992b) A homology domain shared between Drosophila optomotor-blind and mouse Brachyury is involved in DNA binding. Biochem Biophys Res Commun 186:918–925
Poeck B (1992) Untersuchungen zur Genetik und Expression im optomotor-blind (omb) Genlokus von Drosophila melanogaster. Dissertation, Universität Würzburg
Poeck B, Hofbauer A, Pflugfelder G.O. (1993) Expression of the Drosophila optomotor-blind gene transcript in neuronal and glial cells of the developing nervous system. Development, in press
Ponticelli AS, Smith GR (1992) Chromosomal context dependence of a eukaryotic recombinational hot spot. Proc Nag Acad Sci USA 89:227–231
Ptashne M (1988) How eukaryotic transcriptional activators work. Nature 335:683–689
Shenkar R, Shen M, Arnheim N (1991) DNase I-hypersensitive sites and transcription factor-binding motifs within the mouse Eβ meiotic recombination hot spot. Mol Cell Biol 11:1813–1819
Thomas BJ, Rothstein R (1991) Sex, maps and imprinting. Cell 64:1–3
Thompson PE (1959) New mutants. Drosophila Inf Serv 33:99
Washburn T, O'Tousa JE (1992) Nonsense suppression of the major rhodopsin gene of Drosophila. Genetics 130:585–595
Weinzierl R, Axton JM, Ghysen A, Akam M (1987) Ultrabithorax mutations in constant and variable regions of the protein coding sequence. Genes Dev 1:386–397
Zachar Z, Bingham PM (1982) Regulation of white locus expression: the structure of mutant alleles at the white locus of Drosophila melanogaster. Cell 30:529–541
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Poeck, B., Balles, J. & Pflugfelder, G.O. Transcript identification in the optomotor-blind locus of Drosopbila melanogaster by intragenic recombination mapping and PCR-aided sequence analysis of lethal point mutations. Molec. Gen. Genet. 238, 325–332 (1993). https://doi.org/10.1007/BF00291990
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DOI: https://doi.org/10.1007/BF00291990