scholarly journals ORGANIZATION OF THE ROSY LOCUS IN DROSOPHILA MELANOGASTER: FURTHER EVIDENCE IN SUPPORT OF A CIS-ACTING CONTROL ELEMENT ADJACENT TO THE XANTHINE DEHYDROGENASE STRUCTURAL ELEMENT

Genetics ◽  
1979 ◽  
Vol 91 (2) ◽  
pp. 275-293
Author(s):  
M McCarron ◽  
J O'Donnell ◽  
A Chovnick ◽  
B S Bhullar ◽  
J Hewitt ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Control Region ◽  
Large Scale ◽  
Present Report ◽  
Xanthine Dehydrogenase ◽  
Control Element ◽  
Genetic Dissection ◽  
Structural Element ◽  
Cis Acting ◽  
A Site

ABSTRACT The present report summarizes our recent progress in the genetic dissection of an elementary genetic unit in a higher organism, the rosy locus (ry: 3-52.0) in Drosophila melanogaster. Pursuing the hypothesis that the rosy locus includes a noncoding control region, as well as a structural element coding for the xanthine dehydrogenase (XDH) peptide, experiments are described that characterize and map a rosy locus variant associated with much lower than normal levels of XDH activity. Experiments are described that fail to relate this phenotype to alteration in the structure of the XDH peptide, but clearly associate this character with variation in number of molecules of XDH per fly. Large-scale fine-structure recombination experiments locate the genetic basis for this variation in the number of molecules of XDH per fly to a site immediately to the left of the XDH structural element within a region previously designated as the XDH control element. Moreover, experiments clearly separate this "underproducer" variant site from a previously described "overproducer" site within the control region. Examination of enzyme activity in electrophoretic gels of appropriate heterozygous genotypes demonstrates the cis-acting nature of this variation in the number of molecules of XDH. A revision of the map of the rosy locus, structural and control elements is presented in light of the additional mapping data now available.

ORGANIZATION OF THE ROSY LOCUS IN DROSOPHILA MELANOGASTER: EVIDENCE FOR A CONTROL ELEMENT ADJACENT TO THE XANTHINE DEHYDROGENASE STRUCTURAL ELEMENT

Genetics ◽  
1976 ◽  
Vol 84 (2) ◽  
pp. 233-255
Author(s):  
A Chovnick ◽  
W Gelbart ◽  
M McCarron ◽  
B Osmond ◽  
E P M Candido ◽  
...  
Keyword(s):  
Large Scale ◽  
Genetic Basis ◽  
Structural Information ◽  
Natural Populations ◽  
Control Element ◽  
Structural Element ◽  
Electrophoretic Variants ◽  
Cis Acting ◽  
A Site ◽  
Recombination Experiments

ABSTRACT From a collection of electrophoretic variants of XDH obtained from laboratory strains and natural populations, a stock was isolated that was associated with much greater than normal levels of XDH activity. Preliminary recombination experiments demonstrated that this character maps to the rosy locus. While a series of observations failed to relate this phenotype to alteration in the structure of the XDH polypeptide, kinetic and immunological experiments did succeed in associating this character with variation in number of molecules of XDH/fly. Large scale fine structure recombination experiments locate the genetic basis for this variation in number of molecules of XDH/fly to a site very close to, but definitely outside of, the genetic boundaries of the XDH structural information. Observations are described which eliminate the possibility that we are dealing with a tandem duplication of the XDH structural element. Turning to a regulatory role for this genetic element located adjacent to the XDH structural information, a simple experiment is described which demonstrates that it functions as a "cis-acting" regulator of the XDH structural element.

scholarly journals TISSUE-SPECIFIC AND PRETRANSLATIONAL CHARACTER OF VARIANTS OF THE ROSY LOCUS CONTROL ELEMENT IN DROSOPHILA MELANOGASTER

Genetics ◽  
1984 ◽  
Vol 108 (4) ◽  
pp. 953-968
Author(s):  
S H Clark ◽  
S Daniels ◽  
C A Rushlow ◽  
A J Hilliker ◽  
A Chovnick
Keyword(s):  
Drosophila Melanogaster ◽  
Fat Body ◽  
Present Report ◽  
Malpighian Tubules ◽  
Phenotypic Characterization ◽  
Control Element ◽  
Structural Element ◽  
Tissue Specific ◽  
Cis Acting

ABSTRACT Prior reports from this laboratory have described the experimental basis for our understanding of the genetic organization of the rosy locus (ry:3-52.0) of Drosophila melanogaster, as a bipartite genetic entity consisting of a structural element that codes for the xanthine dehydrogenase (XDH) peptide and a contiguous, cis-acting control element. The present report describes our progress in the analysis of the control element and its variants. Characterization of the control element variants reveals that, with respect to late third instar larval tissue distribution of XDH activity and cross-reacting material, i409H is associated with a large, tissue-specific increase in fat body which is not observed in malpighian tubules. Further data are presented in support of the inference that this differential expression must reflect differential production of XDH-specific RNA transcripts.—Gel blot analyses are described which demonstrate (1) that the phenotypic effects associated with variation in the rosy locus control element relate to differences in accumulation of XDH-specific poly-A+ RNA and (2) do not relate to differences in rosy DNA template numbers.—Experiments are described that provide for unambiguous mapping of control element sites through the use of half-tetrad recombination experiments and the recovery and phenotypic characterization of the reciprocal products of exchange between control element site variants. Thus, we are able to order the sites as follows: kar-i1005 i409-ry.

scholarly journals Genetic analysis of the right (3′) end of the rosy locus in Drosophila melanogaster

1986 ◽  
Vol 47 (2) ◽  
pp. 109-116 ◽  
Author(s):  
S. H. Clark ◽  
A. J. Hilliker ◽  
A. Chovnick
Keyword(s):  
Drosophila Melanogaster ◽  
Genetic Analysis ◽  
Xanthine Dehydrogenase ◽  
Control Element ◽  
Experimental Basis ◽  
Novel Mutations ◽  
Structural Element ◽  
Definition Of ◽  
The Right ◽  
Genetic Entity

SUMMARYPrior reports from this laboratory have described the experimental basis for our understanding of the rosy locus (ry: 3–52·0) of Drosophila melanogaster as a bipartite genetic entity consisting of a structural element that codes for the xanthine dehydrogenase (XDH) peptide and a contiguous cis-acting control element immediately to the left of the structural element. Although the left end (5′) of the structural element has been well defined, the right boundary (3′) has been given only casual treatment in our prior reports. In our recent studies of rosy locus expression we have been concerned with the production and identification of mutations in the non-structural regions immediately flanking the structural element. An improved definition of the right end of the structural element is essential to this analysis. In addition to producing a better definition of the right boundary of the structural element, this study produced several phenotypically novel mutations. These mutations were classified initially ascontrol element mutations, but upon analysis were found to map within the rosy structural element. No evidence was obtained for the existence of a control element contiguous with the right end of the structural element.

scholarly journals INTRACISTRONIC MAPPING OF ELECTROPHORETIC SITES IN DROSOPHILA MELANOGASTER: FIDELITY OF INFORMATION TRANSFER BY GENE CONVERSION

Genetics ◽  
1974 ◽  
Vol 76 (2) ◽  
pp. 289-299
Author(s):  
Margaret McCarron ◽  
William Gelbart ◽  
Arthur Chovnick
Keyword(s):  
Drosophila Melanogaster ◽  
Information Transfer ◽  
Large Scale ◽  
Genetic Basis ◽  
Xanthine Dehydrogenase ◽  
Specific Protein ◽  
Wild Type ◽  
Electrophoretic Variation ◽  
The Stability ◽  
Recombination Experiments

ABSTRACT A convenient method is described for the intracistronic mapping of genetic sites responsible for electrophoretic variation of a specific protein in Drosophila melanogaster. A number of wild-type isoalleles of the rosy locus have been isolated which are associated with the production of electrophoretically distinguishable xanthine dehydrogenases. Large-scale recombination experiments were carried out involving null enzyme mutants induced on electrophoretically distinct wild-type isoalleles, the genetic basis for which is followed as a nonselective marker in the cross. Additionally, a large-scale recombination experiment was carried out involving null enzyme rosy mutants induced on the same wild-type isoallele. Examination of the electrophoretic character of crossover and convertant products recovered from the latter experiment revealed that all exhibited the same parental electrophoretic character. In addition to documenting the stability of the xanthine dehydrogenase electrophoretic character, this observation argues against a special mutagenesis hypothesis to explain conversions resulting from allele recombination studies.

scholarly journals The rosy locus in Drosophila melanogaster: xanthine dehydrogenase and eye pigments.

Genetics ◽  
1991 ◽  
Vol 129 (4) ◽  
pp. 1099-1109 ◽  
Author(s):  
A G Reaume ◽  
D A Knecht ◽  
A Chovnick
Keyword(s):  
Drosophila Melanogaster ◽  
Structural Integrity ◽  
Present Report ◽  
Specific Interaction ◽  
Pigment Granule ◽  
Ultrastructural Localization ◽  
Xanthine Dehydrogenase ◽  
Pigment Formation ◽  
Antibody Staining ◽  
Pigment Granules

Abstract The rosy gene in Drosophila melanogaster codes for the enzyme xanthine dehydrogenase (XDH). Mutants that have no enzyme activity are characterized by a brownish eye color phenotype reflecting a deficiency in the red eye pigment. Xanthine dehydrogenase is not synthesized in the eye, but rather is transported there. The present report describes the ultrastructural localization of XDH in the Drosophila eye. Three lines of evidence are presented demonstrating that XDH is sequestered within specific vacuoles, the type II pigment granules. Histochemical and antibody staining of frozen sections, as well as thin layer chromatography studies of several adult genotypes serve to examine some of the factors and genic interactions that may be involved in transport of XDH, and in eye pigment formation. While a specific function for XDH in the synthesis of the red, pteridine eye pigments remains unknown, these studies present evidence that: (1) the incorporation of XDH into the pigment granules requires specific interaction between a normal XDH molecule and one or more transport proteins; (2) the structural integrity of the pigment granule itself is dependent upon the presence of a normal balance of eye pigments, a notion advanced earlier.

scholarly journals GENETIC ANALYSIS OF THE CENTROMERIC HETEROCHROMATIN OF CHROMOSOME 2 OF DROSOPHILA MELANOGASTER: DEFICIENCY MAPPING OF EMS-INDUCED LETHAL COMPLEMENTATION GROUPS

Genetics ◽  
1976 ◽  
Vol 83 (4) ◽  
pp. 765-782
Author(s):  
Arthur J Hilliker
Keyword(s):  
Drosophila Melanogaster ◽  
Present Report ◽  
Heterochromatic Region ◽  
Centromeric Heterochromatin ◽  
Genetic Constitution ◽  
Genetic Dissection ◽  
Chromosome 2 ◽  
Multiple Alleles ◽  
Products Analysis ◽  
Dominance Tests

ABSTRACT Until recently, little was known of the genetic constitution of the heterochromatic segments of the major autosomes of Drosophila melanogaster. Our previous report described the genetic dissection of the proximal, heterochromatic region of chromosome 2 of Drosophila melanogasterby means of a series of overlapping deficiencies generated by the detachment of compound second autosomes (Hilliker and Holm 1975). Analysis of these deficiencies by inter se complementation, pseudo-dominance tests with proximal mutations and allelism tests with known deficiencies provided evidence for the existence of at least two loci between the centromere and the light locus in 2L and one locus in 2R between the rolled locus and the centromere. These data in conjunction with cytological observations demonstrated that light and rolled and three loci lying between them are located within the proximal heterochromatin of the second chromosome.——The present report describes the further analysis of this region through the induction with ethyl methanesulphonate (EMS) of recessive lethals allelic to the 2L and 2R proximal deficiencies associated with the detachment products. Analysis of the 118 EMS-induced recessive lethals and visible mutations recovered provided evidence for seven loci in the 2L heterochromatin and six loci in the 2R heterochromatin, with multiple alleles being obtained for most sites. Of these loci, one in 2L and two in 2R fall near the heterochromatic-euchromatic junctions of 2L and 2R respectively. None of the 113 EMS lethals behaved as a deficiency, implying that the heterochromatic loci uncovered in this study represent nonrepetitive cistrons. Thus functional genetic loci are found in heterochromatin, albeit at a very low density relative to euchromatin.

ON THE IDENTIFICATION OF THE ROSY LOCUS DNA IN DROSOPHILA MELANOGASTER: INTRAGENIC RECOMBINATION MAPPING OF MUTATIONS ASSOCIATED WITH INSERTIONS AND DELETIONS

Genetics ◽  
1986 ◽  
Vol 112 (4) ◽  
pp. 755-767
Author(s):  
S H Clark ◽  
M McCarron ◽  
C Love ◽  
A Chovnick
Keyword(s):  
Drosophila Melanogaster ◽  
Large Scale ◽  
Conclusive Evidence ◽  
Hybrid Dysgenesis ◽  
Intragenic Recombination ◽  
Wild Type ◽  
Structural Element ◽  
Insertions And Deletions ◽  
Recombination Mapping

ABSTRACT DNA extracts of several rosy-mutation-bearing strains were associated with large insertions and deletions in a defined region of the molecular map believed to include the rosy locus DNA. Large-scale, intragenic mapping experiments were carried out that localized these mutations within the boundaries of the previously defined rosy locus structural element. Molecular characterization of the wild-type recombinants provides conclusive evidence that the rosy locus DNA is localized to the DNA segment marked by these lesions.—One of the mutations, ry  2101, arose from a P-M hybrid dysgenesis experiment and is associated with a copia insertion. Experiments are described which suggest that copia mobilizes in response to P-M hybrid dysgenesis.—Relevance of the data to recombination in higher organisms is considered.

scholarly journals Mutations Affecting Expression of the rosy Locus in Drosophila melanogaster

Genetics ◽  
1987 ◽  
Vol 116 (1) ◽  
pp. 55-66
Author(s):  
Chong Sung Lee ◽  
Daniel Curtis ◽  
Margaret McCarron ◽  
Carol Love ◽  
Mark Gray ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Xanthine Dehydrogenase ◽  
Control Element ◽  
Regulatory Change ◽  
Cdna Clones ◽  
Cdna Sequences ◽  
Regulatory Mutations ◽  
New Mutations

ABSTRACT The rosy locus in Drosophila melanogaster codes for the enzyme xanthine dehydrogenase (XDH). Previous studies defined a "control element" near the 5′ end of the gene, where variant sites affected the amount of rosy mRNA and protein produced. We have determined the DNA sequence of this region from both genomic and cDNA clones, and from the ry  +10 underproducer strain. This variant strain had many sequence differences, so that the site of the regulatory change could not be fixed. A mutagenesis was also undertaken to isolate new regulatory mutations. We induced 376 new mutations with 1-ethyl-1-nitrosourea (ENU) and screened them to isolate those that reduced the amount of XDH protein produced, but did not change the properties of the enzyme. Genetic mapping was used to find mutations located near the 5′ end of the gene. DNA from each of seven mutants was cloned and sequenced through the 5′ region. Mutant base changes were identified in all seven; they appear to affect splicing and translation of the rosy mRNA. In a related study (T. P. Keith et al. 1987), the genomic and cDNA sequences are extended through the 3′ end of the gene; the combined sequences define the processing pattern of the rosy transcript and predict the amino acid sequence of XDH.

Observations on chromosomal male sterility in Drosophila melanogaster

1984 ◽  
Vol 26 (1) ◽  
pp. 67-77
Author(s):  
James. C. Stone
Keyword(s):  
Drosophila Melanogaster ◽  
X Chromosome ◽  
Functional Unit ◽  
Primary Spermatocyte ◽  
Male Meiosis ◽  
Centromeric Heterochromatin ◽  
Control Element ◽  
Male Sterile ◽  
Cis Acting ◽  
Autosome Translocation

Observations on a variety of metazoans have shown that the X chromosome becomes functionally inactive earlier in male meiosis than the remainder of the genome. Genetic analyses of male-sterile chromosome rearrangements in Drosophila suggest that the X chromosome in this species behaves as a distinct functional unit, and have further suggested that X-chromosome expression is regulated in the primary spermatocyte by a cis-acting control element located in the centromeric heterochromatin. Attempts to test the X-inactivation hypothesis of chromosomal sterility in Drosophila and attempts to map the hypothetical control element are described here. Cytological observations on a male-sterile X-autosome translocation are also discussed.

GENETIC LIMITS OF THE XANTHINE DEHYDROGENASE STRUCTURAL ELEMENT WITHIN THE ROSY LOCUS IN DROSOPHILA MELANOGASTER

Genetics ◽  
1974 ◽  
Vol 78 (3) ◽  
pp. 869-886
Author(s):  
William M Gelbart ◽  
Margaret McCarron ◽  
Janardan Pandey ◽  
Arthur Chovnick
Keyword(s):  
Drosophila Melanogaster ◽  
Amino Acid ◽  
Structural Information ◽  
Xanthine Dehydrogenase ◽  
Gene Organization ◽  
Null Mutant ◽  
Electrophoretic Variation ◽  
Structural Element ◽  
Electrophoretic Variants ◽  
Left And Right

Abstract Experiments are described that provide an opportunity to estimate the genetic limits of the structural (amino acid coding) portion of the rosy locus (3: 52.0) in Drosophila melanogaster, which controls the enzyme, xanthine dehydrogenase (XDH) . This is accomplished by mapping experiments which localize sites responsible for electrophoretic variation in the enzyme on the known genetic map of null-XDH rosy mutants. Electrophoretic sites are distributed along a large portion of the null mutant map. A cis-trans test involving electrophoretic variants in the left- and right-hand portions of the map leads to the conclusion that the entire region between these variants is also structural. Hence most, if not all, of the null mutant map of the rosy locus contains structural information for the amino acid sequence of the XDH polypeptide. Consideration is given to the significance of the present results for the general problem of gene organization in higher eukaryotes.

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