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  • 1
    ISSN: 1573-4927
    Keywords: l-glycerol-3-phosphate dehydrogenase (α-GPDH) ; isozymes ; development ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The basis for the differentiation of l-glycerol-3-phosphate dehydrogenase (α-GPDH) into larval and adult isozymes in Drosophila melanogaster was investigated by the correlation of a lack of appearance of each isozyme during development within Drosophila bearing α-GPDH “null” alleles and by the study of a putative conversion factor. Conversion studies indicate the presence of a heat-labile RNase-resistant conversion factor present in crude larval extracts with the ability to convert GPDH-1 to GPDH-2 and GPDH-3 but not vice versa. In addition, “null” mutations at the Gpdh locus obliterate all isozymatic species of α-GPDH in all developmental stages. These observations suggest that all α-GPDH isozymes are the product of a single structural gene and that the multiple forms of this enzyme arise during successive developmental stages through an epigenetic modification of the primary Gpdh + polypeptide. Finally, observations are reported which bear on the functional divergence of the α-glycerophosphate cycle in the adult and larval stage of development.
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  • 2
    ISSN: 1573-4927
    Keywords: α-GPD null mutants ; immunoelectrophoresis ; two-dimensional electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A protein spot corresponding to l-glycerol-3-phosphate dehydrogenase (α-GPDH, E.C. 1.1.1.8, NAD+ oxidoreductase) has been identified on a two-dimensional gel (isoelectric focusing-SDS gel) containing up to 150 stained protein spots from a crude Drosophila homogenate. Preliminary identification of the α-GPDH spot was made by including a suitable amount of purified Drosophila α-GPDH in crude fly homogenates prior to electrophoresis and observing an intensity enhancement of the corresponding protein spot on the gels. When three purified electrophoretic variants (slow, fast, and ultrafast) were mixed and analyzed by two-dimensional gel electrophoresis, horizontal displacements of the three protein spots were observed. Immunoprecipitation of the enzyme prior to electrophoresis and gene mapping further confirmed the identity of the α-GPDH protein spot. The α-GPDH spot can also be detected by autoradiography of a two-dimensional gel from a single fly extract, where it has been estimated to constitute 0.5–1% of the total soluble protein. Mutants which express no apparent α-GPDH activity were analyzed by two-dimensional gels and immunoelectrophoresis in an attempt to identify and characterize the inactive proteins. It is suggested that these techniques provide a powerful tool for the analysis of CRM+-null activity mutants of a specific gene-enzyme system.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Biochemical genetics 16 (1978), S. 769-775 
    ISSN: 1573-4927
    Keywords: hidden variation ; α-GPDH ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The level of hidden variation in populations of Drosophila melanogaster at the Gpdh + locus was determined by thermal stability studies of the protein. The results indicate a lack of variation using these methods both in and between the two common electrophoretic variants. It is suggested that α-GPDH is conserved in primary structure, which may be related to its critical role in flight muscle metabolism.
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  • 4
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Null and low activity alleles at the genetic locus coding for L-Glycerol-3-phosphate dehydrogenase (α-GPDH, NAD+ oxidoreductase, E.C. 1.1.1.8) in Drosophila melanogaster have been analyzed by a combination of rocket immunoelectrophoresis, interallelic complementation, and two-dimensional gel electrophoresis. In addition to providing information on the molecular weight, charged state, and steady state level of CRM in each of these mutants, it is suggested that each mutation has resulted in a genetic lesion within the structural element, Gpdh +. CRM levels appear to be the result of a differential sensitivity to the normal intracellular degradative process and the CRM- mutants represent “hypersensitive” alleles, such that the mutant polypeptide does not accumulate in the intracellular environment.
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  • 5
    ISSN: 0192-253X
    Keywords: catalase ; Drosophila ; development ; turnover ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ontogenetic and tissue-specific expression of catalase (E.C. 1.11.1.6) has been determined in a wild type strain of Drosophila melanogaster derived from a natural population. Two distinct peaks of activity are observed during development with the first peak occurring in late third instar larvae just prior to puparium formation, and the second and larger of the two peaks occurring during metamorphosis. These peaks of catalase activity are coincident with the two major peaks of ecdysone titer. Of the tissues assayed, larval malpighian tubules, gut, and fat body demonstrated the highest specific activities. Adult abdomen exhibited a two- to three-fold higher specific activity than either head or thorax. Of the abdominal tissues assayed, malpighian tubules and abdominal wall had the highest specific activities. Malpighian tubules were the only sexually dimorphic tissue with respect to catalase activity and are apparently largely responsible for an overall increase observed in female abdominal activity. Catalase-specific CRM levels parallel the enzyme activity levels indicating that these tissue-specific activity differences reflect differences in the rate of accumulation of catalase molecules.Turnover studies employing the catalase inhibitor 3-amino-1,2,4-triazole were conducted on head, thorax, and abdomen of male adult flies. Rates of catalase degradation were similar in the three body segments with a slightly higher rate in abdominal tissue. Therefore the different steady state levels observed largely reflect different rates of catalase synthesis.
    Additional Material: 5 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 3 (1982), S. 129-142 
    ISSN: 0192-253X
    Keywords: glycerol phosphate dehydrogenase ; turnover ; Drosophila ; gene dosage ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In vivo radiolabeling of Drosophila melanogaster sn-glycerol-3-phosphate dehydrogenase (E.C. 1.1.1.8; GPDH) has been accomplished by microinjection of 3H-leucine into anesthetized flies. Comigration of immunoprecipitated radiolabeled GPDH with purified 14C-labeled GPDH-1 in SDS polyacrylamide disc gels has established the monospecificity of our immunoprecipitation technique. Short-term uptake experiments have demonstrated that maximum radiolabel incorporation of total TCA precipitable protein and immunoprecipitable GPDH-1 occurs within 4 hours postinjection, with GPDH-1 accounting for approximately 1% of the total radiolabeled TCA precipitable protein. In order to develop the parameters for turnover studies of GPDH in Drosophila, a comparative analysis of the rates of synthesis and degradation of GPDH-1 in flies bearing two and three doses of the structural gene have been conducted by the construction of adult flies aneuploid and euploid for the cytogenetic region 25F-26B on the left arm of chromosome II. Short-term uptake studies have demonstrated that the rate of GPDH-1 synthesis in the three-dose flies is approximately 1.58 times that found in the two-dose euploid flies. This value is in close agreement with data obtained for steady-state levels of CRM by rocket immunoelectrophoresis. In contrast, longterm pulse-chase experiments have revealed that rates of GPDH-1 degradation in these aneumploid and euploid flies appear to be identical. These data suggest that the rate of GPDH-1 synthesis in Drosophila is primarily regulated by a tightly linked cis-acting element which appears to act autonomously with respect to gene copy number as well as steady-state GPDH protein levels.
    Additional Material: 5 Ill.
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  • 7
    ISSN: 0192-253X
    Keywords: temporal genes ; GPDH isozymes ; regulation ; development, Drosophila ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The complete developmental program of glycerol-3-phosphate dehydrogenase in wild type Drosophila is described with respect to activity, isozyme expression, and GPDH-specific CRM. Variants of this developmental program have been isolated from natural populations which affect the rate of accumulation of only the GPDH-3 isozyme in both the larval and adult stages of development. This activity variation segregates as a single gene which is tightly linked to the structural element on Chromosome II, exhibits cis-control, and is tissue specific in expression. This gene meets all the criteria for temporal regulatory genes.
    Additional Material: 8 Ill.
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  • 8
    ISSN: 0192-253X
    Keywords: Glycerol-3-phosphate dehydrogenase ; tissue-specific expression ; isozymic composition ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The tissue-specific expression and isozymic composition of Drosophila sn-glycerol-3-phosphate dehydrogenase (GPDH) (EC 1.1.1.8) have been determined for a high-activity control line and two variant lines that alter either the temporal or systemic expression of GPDH through a reduction in rates of polypeptide synthesis. The temporal variant exhibits a reduction in enzyme levels in all larval tissues and in the adult abdomen, while levels of activity in the adult thorax are equal to the control line. Isozymic analyses of these tissues demonstrate that it is the GPDH-3 species that is reduced in a temporal and tissue-specific manner. In contrast, the systemic variant demonstrates a uniform reduction of all isozymic species in each tissue and developmental stage. Analyses of the tissues of F1 hybrid offspring of each variant line and appropriately marked electrophoretic variants demonstrate that the tissue-specific effects observed are due to cis-acting elements that are tightly linked to the structural gene.
    Additional Material: 6 Ill.
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  • 9
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 59-60 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 10
    Publication Date: 1980-10-01
    Print ISSN: 0006-2928
    Electronic ISSN: 1573-4927
    Topics: Biology , Chemistry and Pharmacology
    Published by Springer
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