Organization of the rosy locus in Drosophila melanogaster

Chovnick, Arthur; Gelbart, William M.; McCarron, Margaret

doi:10.1016/0092-8674(77)90312-9

Cited by 110 publications

(34 citation statements)

References 25 publications

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“…Genetically these regions appear to be recalcitrant to mutation that is phenotypically visible (32)(33)(34). Molecular studies indicate that independent transcriptional units are often separated by large intergenic regions (31).…”

Section: Discussionmentioning

confidence: 99%

Restriction map variation in the Adh region of Drosophila.

Langley¹,

Montgomery²,

Quattlebaum³

1982

Proc. Natl. Acad. Sci. U.S.A.

105

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Restriction maps of a 12,000-nucleotide region containing the alcohol dehydrogenase (Adh) locus of the second chromosome of Drosophila were determined for (i) 18 independent second chromosome stocks of D. melanogaster established from several natural populations and (ii) five closely related Drosophila species. All of the detected map differences appear to lie outside the Adh coding block. Four polymorphic restriction sites and four insertion/deletion variants were identified among D. melanogaster chromosomes. The estimated heterozygosity per nucleotide site was 0.006. The relative distributions of restriction sites and insertion/deletion variations among subpopulations and species suggest that the insertion/deletion variation may be mildly deleterious.In recent years it has become possible to survey organisms for DNA sequence variation detected as differences in distributions of restriction endonuclease sites along specific regions of DNA (1). Both nuclear and cytoplasmic DNAs show variation in laboratory strains. These differences are important in localizing mutational lesions and functional regions. Restriction map variation also has been systematically surveyed in natural populations of several species, including Drosophila melanogaster, simulans, and virilis (2); the deer mouse (3); and man (4). These studies have allowed the estimation of .sequence diversity among individuals, populations, and species in DNA sequences that have been inaccessible (e.g., noncoding regions and mitochondrial genomes).Goldberg reported the cloning ofthe DNA in and around the alcohol dehydrogenase locus (Adh) of D. melanogaster (5). A single transcript was found from this region-that of Adh (5). The availability of this cloned sequence afforded us the opportunity to survey restriction map variation in this region. We report here the restriction maps from 18 independent second chromosome stocks of D. melanogaster established from several natural populations. We also present restriction maps for five other species closely related to D. melanogaster. From these maps, estimates of sequence diversity over a 12,000-nucleotide region can be made. Insertion/deletion variation was found both within and between species. The distribution over individuals, subpopulations, and species suggests some selective effect of insertions/deletions outside the transcriptional unit.MATERIALS AND METHODS Drosophila Stocks. Eighteen stocks of D. melanogaster homozygous for independent second chromosomes isolated from several natural populations were received from C. C. Laurie-Ahlberg (6). Ten

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Section: Discussionmentioning

confidence: 99%

Restriction map variation in the Adh region of Drosophila.

Langley¹,

Montgomery²,

Quattlebaum³

1982

Proc. Natl. Acad. Sci. U.S.A.

105

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“…melanogaster strains and large-scale growth. The following wild-type D. rnelanogaster strains were used (Gelbart et al, 1974;Chovnick et al, 1976Chovnick et al, , 1977McCarron et al, 1976; abbreviations are given in square brackets): ry+ ' [wt], ry+"' [wt(+10)], ry" [wt(+4)], Canton S [wt(S)]. The last named strain is multi-allelic.…”

Section: Methodsmentioning

confidence: 99%

Properties of Xanthine Dehydrogenase Variants from Rosy Mutant Strains of Drosophila melanogaster and their Relevance to the enzyme's Structure and Mechanism

Doyle

Burke

Chovnick

et al. 1996

European Journal of Biochemistry

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Xanthine dehydrogenase, a molybdenum, iron-sulfur tlavoenzyme encoded in the fruit fly Drosophila melanogaster by the rosy gene, has been characterised both from the wild-type and mutant flies. Enzyme assays, using a variety of different oxidising and reducing substrates were supplemented by limited molecular characterisation. Four rosy strains showed no detectable activity in any enzyme assay tried, whereas from four wild-type and three rosy mutant strains, those for the [E89K], [L127F] and [L157P]xanthine dehydrogenases (in all of which the mutation is in the iron-sulfur domain), the enzyme molecules, although present at different levels, had extremely similar or identical properties. This was confirmed by purification of one wild-type and one mutant enzyme, LE89Klxanthine dehydrogenase. These both had ultraviolet-visible absorption spectra similar to milk xanthine oxidase. Both were found to be quite stable molecules, showing very high catalytic-centre activities and with little tendency to become degraded by proteolysis or modified by conversion to oxidase or desulfo forms. In three further rosy strains, giving [G353D]xanthine dehydrogenase and IS357FIxanthine dehydrogenase mutated in the flavin domain, and [GI 01 IEIxanthine dehydrogenase mutated in the molybdenum domain, enzyme activities were selectively diminished in certain assays. For the G353D and S357F mutant enzymes activities to NAD' as oxidising substrate were diminished, to zero for the latter. In addition for [G353D]xanthine dehydrogenase, there was an increase in apparent K,, values both for NAD ' and NADH. These findings indicate involvement of this part of the sequence in the NAD+-binding site. The G l O l l E mutation has a profound effect on the enzyme. As isolated and as present in crude extracts of the flies, this xanthine dehydrogenase variant lacks activity to xanthine or pterin as reducing substrate, indicating an impairment of the functioning of its molybdenum centre. However, it retains full activity to NADH with dyes as oxidising substrate. Mild oxidation of the enzyme converts it, apparently irreversibly, to a form showing full activity to xanthine and pterin. The nature of the group that is oxidised is discussed in the light of redox potential data. It is proposed that the process involves oxidation of the pterin of the molybdenum cofactor from the tetrahydro to a dihydro oxidation state. This conclusion is fully consistent with recent information [Romiio, M. J., Archer, M., Moura, I., Moura, J. J. G., LeGall, J., Engh, R., Schneider, M., Hof, P. & Huber, R. (1995) Science 270, 1170-11761 from X-ray crystallography on the structure of a closely related enzyme from Desulfovibrio gigas. It is proposed, that apparent irreversibility of the oxidative activating process for [GI 01 IEIxanthine dehydrogenase, is due to conversion of its pterin to the tricyclic derivative detected by these workers. The data thus provide the strongest evidence available, that the oxidation state of the pterin can have a controlling influence on the activit...

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“…From the total thickness of these bands and from preliminary data on recombination frequencies between point mutants in the region (:0. 1 map units), an estimate ofthe DNA contained in Sh would be of the order of 100 kilobases (24). The Sh gene appears to contain three major areas: two flanking regions in which chromosome breaks or mutations result in Sh phenotypes and a central region that, when deleted, results in haplolethality.…”

Section: Methodsmentioning

confidence: 99%

Abnormal action potentials associated with the Shaker complex locus of Drosophila

Tanouye

Ferrús

Fujita

1981

Proc. Natl. Acad. Sci. U.S.A.

154

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Intracellular recordings ofaction potentials were made from the cervical giant axon in Shaker (Sh) mutants and normal Drosophila. The mutants showed abnormally long delays in repolarization. The defect is not due to abnormal Ca2+ channels, because it persists in the presence of Co2+, blocker. On the other hand, the K+-channel blocker 4-aminopyridine causes a similar effect in normal animals, suggesting that the

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Organization of the rosy locus in Drosophila melanogaster

Cited by 110 publications

References 25 publications

Restriction map variation in the Adh region of Drosophila.

Restriction map variation in the Adh region of Drosophila.

Properties of Xanthine Dehydrogenase Variants from Rosy Mutant Strains of Drosophila melanogaster and their Relevance to the enzyme's Structure and Mechanism

Abnormal action potentials associated with the Shaker complex locus of Drosophila

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