Peroxisomes of the firefly lantern

Hanna, C. H.; Hopkins, Thomas A.; Buck, John D.

doi:10.1016/s0022-5320(76)80105-0

Cited by 45 publications

(16 citation statements)

References 27 publications

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“…Among insects, peroxisomes have been identified cytochemically in Calpodes, Gryllus, and Photuris, where peroxisomes were studied in larval oenocytes and fat body and adult lantern (27)(28)(29)(30)(31).…”

Section: Discussionmentioning

confidence: 99%

Peroxisomes in wild-type and rosy mutant Drosophila melanogaster.

Beard

Holtzman

1987

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

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This study shows that peroxisomes are abundant in the Malpighian tubule and gut of wild-type Oregon R Drosophila melanogaster and that the peroxisomal population of the rosy-506 eye-color mutant differs from that of the wild type. Catalase activity in wild-type flies is demonstrable in bodies of appearance and centrifugal behavior comparable to the perixosomes of vertebrate tissues. Xanthine oxidase (xanthine:oxygen oxidoreductase, EC 1. Deficits in one or more of the peroxisomal enzymes and possible defects in peroxisomal structure are correlated with certain human inherited metabolic diseases (1-4). These diseases are still poorly understood, in part because different tissues show different effects of the mutation and in part because alterations in single genes can seemingly have multiple effects on the peroxisomes. Studies of the disorders are made difficult by their rareness and by the obvious limits of working with human material. It would be helpful to study analogues of the disorders in animals amenable to detailed analysis, but thus far such analogues have not been available. We chose to initiate our explorations for peroxisomal mutants in Drosophila by comparing wild-type Oregon R strain flies with the rosy-506 eye-color mutant, in which 90% of the structural gene for xanthine dehydrogenase (xanthine: NAD+ oxidoreductase, EC 1.1.1.204) is deleted (5, 6). This protein is a bifunctional oxidoreductase that can act as a dehydrogenase or as an oxidase (xanthine:oxygen oxidoreductase, EC 1.1.3.22) depending upon substrate and acceptor availability (7,8). The oxidase activity of the enzyme is critical to the degradation of purines in many organisms; the dehydrogenase function is central to the formation of pterinebased eye-color pigments in Drosophila. We suspected that this protein might be peroxisomal because (i) when acting as an oxidase, it produces hydrogen peroxide, a common feature of peroxisomal oxidases; (ii) it requires flavin in generating peroxide, as is found for many other peroxisomal oxidases; and (iii) its role in punine degradation is potentially functionally linked to activity of allantoicase and uricase, known peroxisomal enzymes (9). The subcellular localization of xanthine oxidase is not clearly established. Some cell fractionation studies show most of the enzyme to be soluble (9, 10), while other investigations report some xanthine oxidase cosedimenting with peroxisomal enzymes (11, 12).The study reported here identifies peroxisomes in the Malpighian tubule and gut of adult Drosophila wild-type Oregon R and rosy-506 strains, by virtue of their content of catalase, the peroxisomal "marker" enzyme. To localize xanthine oxidase, we have adapted cytochemical methods used to demonstrate other oxidases. With these methods, we have shown the presence of xanthine oxidase in peroxisomes of wild-type flies and the absence of this enzyme in rosy-506 flies. We also find signs that, as in the human mutations, a relatively simple genetic change can have complex effects on the peroxisomes. Prel...

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Section: Discussionmentioning

confidence: 99%

Peroxisomes in wild-type and rosy mutant Drosophila melanogaster.

Beard

Holtzman

1987

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

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“…The firefly lantern is comprised of specialized cells called photocytes, which contain large amounts of luciferase (11). Within the photocytes are large numbers of diaminobenzidine-positive granules known as photocyte granules (16). Immunoelectron microscopy experiments performed previously on thin plastic sections of photocytes from Photuris sp., a closely related species, had indicated that luciferase was localized to the photocyte granules (17).…”

Section: Localization Of Luciferase In the Firefly Lantern Usingmentioning

confidence: 99%

Firefly luciferase is targeted to peroxisomes in mammalian cells.

Keller¹,

Gould²,

DeLuca³

et al. 1987

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

171

103

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Although several enzymes known to reside in peroxisomes have been studied extensively, no cis-acting amino acid sequences involved in the transport of these proteins to peroxisomes have been described. As a first step towards the determination of a putative peroxisomal targeting sequence, we have expressed the cDNA encoding the firefly luciferase [Pholinus-luciferin:oxygen 4-oxidoreductase (decarboxylating, ATP-hydrolyzing), EC 1.13.12.7] in monkey kidney cells and found that the product of the gene is transported to peroxisomes. Luciferase is derived from the firefly (Photinus pyralis) and is synthesized and stored in the cells of the firefly's lantern organ, where it is also found in peroxisomes. The fact that this protein is similarly targeted in cells from such different organisms suggests that the process of protein transport to peroxisomes has been highly conserved through evolution.The eukaryotic cell contains distinct organelles, each highly specialized for its particular functions. To maintain this organization, the cell must efficiently direct proteins to their proper subcellular locations. Previous studies dealing with the sorting of proteins to subcellular compartments have demonstrated the necessity of specific sequences or protein modifications for the transport ofproteins to the endoplasmic reticulum (1, 2), lysosomes (3), chloroplasts (4), mitochondria (5), and the nucleus (6-8). Therefore, it is reasonable to assume that analogous cis-acting sequences are involved in the transport of peroxisomal proteins to peroxisomes. At present, virtually nothing is known about the signals that sort peroxisomal proteins. We recently expressed the cloned firefly luciferase gene in CV-1 cells, a monkey kidney cell line. In transfected cells, the gene product was shown by indirect immunofluorescence to be present in small vesicular structures (9). In this paper, we show by double-immunofluorescence experiments that firefly luciferase [Photinus-luciferin:oxygen 4-oxidoreductase (decarboxylating, ATP-hydrolyzing), EC 1.13.12.7] is transported into the peroxisomes of transfected mammalian cells. We also demonstrate by immunocryoelectron microscopy that luciferase is localized within peroxisomes in the cells of the firefly lantern. Since luciferase is not endogenous to mammalian cells, expression of firefly luciferase and altered derivatives of the gene in mammalian cells should provide a valuable model system for future studies on the transport and uptake of proteins into peroxisomes. MATERIALS AND METHODSVectors. The plasmid pRSVL, whose construction is described elsewhere (9), contains the full-length luciferase cDNA under the transcriptional control of the promoter from the Rous sarcoma virus (RSV) long terminal repeat (Fig. 1). At the 3' end of the luciferase gene are the splicing and polyadenylylation sequences from the simian virus 40 (SV40) early region.Transfections. CV-1 monkey kidney cells were plated onto coverslips placed in 10-cm dishes; 24 hr later, the cells were transfected by the calcium ph...

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“…With the electron microscope, they were beautifully analyzed by Smith (1963) in Photuris (adult males). Additional data were given for the same genus by Hanna et al (1976) and Ghiradella (1977;1978), and for adults belonging to other genera by Peterson and Buck (1968) and Ghiradella (1978). In contrast, in Photuris larvae, which glow continuously, the tracheal end organs are not present (Peterson, 1970).…”

Section: Tracheal Cells and Tracheoblastsmentioning

confidence: 99%