Structure and Regulated Expression of the δ-Aminolevulinate Synthase Gene from Drosophila melanogaster

Mena, I; Fernández-Moreno, Miguel Ángel; Börnstein, Belén; Kaguni, Laurie S.; Garesse, Rafael

doi:10.1074/jbc.274.52.37321

Cited by 29 publications

(16 citation statements)

References 62 publications

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“…Localization of the transcriptional start site identified a unique position 22 bp upstream from the initiator methionine codon in adults and multiple start sites in embryos. This difference in transcriptional initiation site usage was also observed in the gene encoding the rate-limiting heme biosynthetic enzyme, ␦-aminolevulinate synthase (43). In both cases, the proximal promoter lacks typical TATA and CAAT boxes in canonical positions.…”

Section: Discussionmentioning

confidence: 79%

Regulation of Mitochondrial Single-stranded DNA-binding Protein Gene Expression Links Nuclear and Mitochondrial DNA Replication inDrosophila

Mena

Lefai

Garesse

et al. 2000

Journal of Biological Chemistry

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The structural organization of the Drosophila melanogaster gene encoding mitochondrial single-stranded DNA-binding protein (mtSSB) has been determined and its pattern of expression evaluated during Drosophila development. The D. melanogaster mtSSB gene contains four exons and three small introns. The transcriptional initiation site is located 22 nucleotides upstream from the initiator translation codon in adults, whereas several initiation sites are found in embryos. No consensus TATA or CAAT sequences are located at canonical positions, although an AT-rich sequence was identified flanking the major transcriptional initiation site. Northern analyses indicated that the mtSSB transcript is present at variable levels throughout development. In situ hybridization analysis shows that maternally deposited mtSSB mRNA is distributed homogeneously in the early embryo, whereas de novo transcript is produced specifically at an elevated level in the developing midgut. Transfection assays in cultured Schneider cells with promoter region deletion constructs revealed that the proximal 230 nucleotides contain cis-acting elements required for efficient gene expression. Putative transcription factor binding sites clustered within this region include two Drosophila DNA replication-related elements (DRE) and a single putative E2F binding site. Deletion and base substitution mutagenesis of the DRE sites demonstrated that they are required for efficient promoter activity, and gel electrophoretic mobility shift analyses showed that DRE binding factor (DREF) binds to these sites. Our data suggest strongly that the Drosophila mtSSB gene is regulated by the DRE/DREF system. This finding represents a first link between nuclear and mitochondrial DNA replication.Animal mitochondria are essential energy-producing organelles that contain multiple copies of their small doublestranded DNA genome (1). Despite its limited coding capacity, mtDNA is critical because it encodes 13 polypeptides that are essential components of mitochondrial respiratory complexes. Biogenesis of mitochondria requires expression and duplication of the mtDNA genome and relies heavily on the nuclear genome, which provides all of the protein components required for these processes and also those involved in their regulation. Mitochondrial DNA polymerase (DNA polymerase ␥) and mitochondrial single-stranded DNA-binding protein (mtSSB) 1 are key nuclear encoded components of the mtDNA replication apparatus: DNA polymerase ␥ is the replicative DNA polymerase (2-4), and mtSSB functions in helix destabilization (5, 6) and in enhancing both the activity and processivity of DNA polymerase ␥ (7, 8). In vivo, mtSSB is associated with the mitochondrial nucleoid (9) and is concentrated within the perinuclear mitochondria that constitute active sites of mtDNA replication (10, 11). Its role is critical in replication because deletion of the yeast protein causes loss of mtDNA (12). mtSSB shares both structural and functional similarities with Escherichia coli SSB (13,14). Both are homo...

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Section: Discussionmentioning

confidence: 79%

Regulation of Mitochondrial Single-stranded DNA-binding Protein Gene Expression Links Nuclear and Mitochondrial DNA Replication inDrosophila

Mena

Lefai

Garesse

et al. 2000

Journal of Biological Chemistry

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“…In embryos, catalase expression parallels closely the expression of d-aminolevulinate synthase (ALAS), the first enzyme in the heme biosynthetic pathway (Ruiz de Mena et al, 1999). Heme serves as a redox prosthetic group of respiratory cytochromes and other hemoproteins, including oxygen carrier proteins.…”

Section: Discussionmentioning

confidence: 98%

Profiling catalase gene expression in Drosophila melanogaster during development and aging

Klichko

Radyuk

Orr

2004

Arch Insect Biochem Physiol

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Catalase represents one of the key antioxidant enzymes (AOE) in the metabolism of oxygen free radicals. A comprehensive analysis was brought to bear on establishing catalase gene expression profiles during development and aging, with the underlying objective being to identify potential regulatory factors. Expression of the catalase gene exhibits substantial variations during development and aging in a stage- and tissue-specific manner. At the temporal level, previous observations of the coincidence of ecdysteroid pulses with peaks in catalase expression during developmental stages were largely corroborated. In adults, a small but significant decline in catalase expression was noted in adults as a function of age. Spatially, it was ascertained that catalase expression is mostly confined to tissues related to intermediary metabolism, digestive and adipose systems as well as oenocytes. By combining histochemical analysis of reporter gene expression with immunostaining of the endogenous product, it was possible to identify putative positive and negative regulatory elements that control catalase expression. Finally, when adult flies were subjected to various environmental insults, such as heat, paraquat, hyperoxia and H(2)O(2), no significant responses were observed, suggesting that catalase gene expression is largely governed by intrinsic genetic programs.

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“…Alas is involved in the synthesis of hemoproteins. The Alas promoter region has been shown to possess regulatory regions that interact with free heme to regulate Alas transcription (Ruiz de Mena et al, 1999). Therefore, it can be hypothesized that under conditions of free heme availability, DmFMO-1 could be transcribed in a truncated form (which would be possible if exons 4 and 5 were not spliced).…”

Section: September 2004mentioning

confidence: 99%

Catalytic activity and expression of two flavin‐containing monooxygenases from Drosophila melanogaster

Scharf

Bennett

et al. 2004

Arch Insect Biochem Physiol

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Two flavin-containing monooxygenase genes occur in the Drosophila genome (named DmFMO-1 and DmFMO-2). Differences exist between these two FMOs in: (1) genomic DNA architecture and predicted post-translational modifications; (2) recombinant protein solubility, activity, and absorbance spectra; and (3) subcellular distribution and developmental transcription/translation profiles in wildtype flies. Characteristic FAD absorbance spectra and strong catalytic competence in methimazole sulfoxidation were observed for recombinant DmFMO-2. Alternatively, weak sulfoxidation was observed for DmFMO-1, which correlated with reduced solubility in the recombinant system. Western blot analyses using specific antisera raised to each FMO showed the two FMOs to be immunologically distinct. In addition, Western blot analyses revealed FMO protein expression in both the microsomal and cytosolic sub-cellular fractions. Interestingly, a larger form of DmFMO-1 occurs in the cytosol that is most strongly expressed in the adult head. These findings suggest divergent physiological roles for DmFMO-1 and DmFMO-2. More specifically, it appears that DmFMO-1 has a distinct developmental role, while DmFMO-2 may have a general housekeeping function.

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Structure and Regulated Expression of the δ-Aminolevulinate Synthase Gene from Drosophila melanogaster

Cited by 29 publications

References 62 publications

Regulation of Mitochondrial Single-stranded DNA-binding Protein Gene Expression Links Nuclear and Mitochondrial DNA Replication inDrosophila

Regulation of Mitochondrial Single-stranded DNA-binding Protein Gene Expression Links Nuclear and Mitochondrial DNA Replication inDrosophila

Profiling catalase gene expression in Drosophila melanogaster during development and aging

Catalytic activity and expression of two flavin‐containing monooxygenases from Drosophila melanogaster

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