Biogenesis of Peroxisomal Proteins in Vivo and in Vitro*

Lazarow, Paul B.; Robbi, Mariette; Fujiki, Yukio; Wong, Laurence

doi:10.1111/j.1749-6632.1982.tb21423.x

Cited by 85 publications

(46 citation statements)

References 13 publications

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“…More speculatively, there could be metabolic links through which the xanthine oxidase enzyme might affect catalase. Some authors posit involvement of xanthine dehydrogenase in iron metabolism, which could influence the availability of heme groups for catalase assembly (8,34). Or, despite the fact that the rosy-506 mutant is derived from the same background stock as the wild type, it may be that genetic effects in the rosy strain influence catalase directly via presently unknown paths.…”

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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“…In rat liver, it is known that matrix proteins in peroxisomes are all synthesized on free polysomes in the cytosol. Post-translational import of these proteins into peroxisomes has been shown by in vivo pulse-chase (6,7) and also by in vitro import experiments (8 -12). We developed an in vitro system based on the import of radiolabeled acyl-CoA oxidase (AOx) 1 into purified rat liver peroxisomes and showed that ATP hydrolysis was required for the translocation of AOx through peroxisomal membranes (9).…”

mentioning

confidence: 99%

Insertion of the 70-kDa Peroxisomal Membrane Protein into Peroxisomal Membranes in Vivo and in Vitro

Imanaka

Shiina

Takano

et al. 1996

Journal of Biological Chemistry

117

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S-PMP70s, with a molecular mass greater than 50 kDa, bound to and inserted into peroxisomal membranes, whereas truncated 35 S-PMP70s smaller than 45 kDa did not. These results suggest that PMP70 is post-translationally transported to peroxisomes without processing and inserted into peroxisomal membranes by a specific mechanism in which a proteinaceous receptor and a certain internal sequence of PMP70 are involved.Peroxisomes are organelles bounded by a single membrane which are present in almost all eukaryotic cells. The peroxisomes are involved in a variety of metabolic processes including peroxide-based respiration, oxidative degradation of purines and fatty acids, and synthesis of plasmalogen and bile acids (1, 2). It has recently been suggested that peroxisomes are formed by post-translational import of newly synthesized proteins into pre-existing peroxisomes, which then divide (3-5). In rat liver, it is known that matrix proteins in peroxisomes are all synthesized on free polysomes in the cytosol. Post-translational import of these proteins into peroxisomes has been shown by in vivo pulse-chase (6, 7) and also by in vitro import experiments (8 -12). We developed an in vitro system based on the import of radiolabeled acyl-CoA oxidase (AOx) 1 into purified rat liver peroxisomes and showed that ATP hydrolysis was required for the translocation of AOx through peroxisomal membranes (9). At least two types of peroxisomal targeting sequences (PTSs) have been found. PTS1 consists of the sequence Ser-Lys-Leu (or closely related) at the carboxyl terminus (10, 13) and PTS2, which in 3-ketoacyl-CoA thiolase is found in the 11 amino acids of the amino-terminal leader sequence (14,15).The membranes of rat liver peroxisomes contain several integral membrane proteins not found in other organelles. Polypeptides of 69/70 and 22 kDa have been identified as major components and polypeptides of 57, 53, 35/36, and 26/27 kDa have been identified as minor components of peroxisomal membranes (16 -19). The 70-kDa peroxisomal membrane protein (PMP70) is markedly induced by administration of hypolipidemic agents and parallels peroxisome proliferation (17, 18). We cloned and sequenced PMP70 and found that PMP70 belonged to a superfamily called an ATP-binding cassette protein, conferring multidrug resistance to tumor cells (20). Recently PMP70 was suggested to be essential for peroxisome formation (21). PMP22 was also cloned and sequenced and it was suggested that the topology of PMP22 was similar to those of Mpv 17 and certain transmitter gated ion channels, although the function of PMP22 has not yet been described (22). Recently peroxisome assembly factor 1, a peroxisomal integral membrane protein with molecular mass of 35 kDa was also cloned and sequenced (23).A number of studies have been carried out to further investigate the biogenesis of peroxisomal membrane proteins (PMPs). We, and other laboratories, have shown that PMP70, -26, and -22 were synthesized on free polysomes (24 -26), suggesting that the PMPs are likely to be insert...

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“…Peroxisome is a ubiquitous organelle found in a wide variety of eukaryotic cells (1). It is a round or oval-shaped structure bounded by a single membrane, ranging from 0.1 to 1.0-m in diameter.…”

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confidence: 99%

“…In humans, deficiency of one of the PEX genes causes peroxisome biogenesis disorders (PBDs), 1 such as Zellweger syndrome, neonatal adrenoleukodystrophy, and infantile Refsum disease (8,9). Patients of Zellweger syndrome lack functional peroxisomes and exhibit severe symptoms in various organs including the central nervous system, and death usually occurs in early childhood.…”

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confidence: 99%

Formation of Peroxisomes from Peroxisomal Ghosts in a Peroxisome-deficient Mammalian Cell Mutant upon Complementation by Protein Microinjection

Yamasaki

Hashiguchi

Fujiwara

et al. 1999

Journal of Biological Chemistry

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Most mammalian cell strains genetically deficient in peroxisome biogenesis have abnormal membrane structures called ghosts, containing integral peroxisomal membrane protein, PMP70, but lacking the peroxisomal matrix proteins. Upon genetic complementation, these mutants regain the ability of peroxisome biogenesis. It is postulated that, in this process, the ghosts act as the precursors of peroxisomes, but there has been no evidence to support this. In the present study, we investigated this issue by protein microinjection to a mutant Chinese hamster ovary cell line defective of PEX5, encoding a peroxisome-targeting signal receptor. When recombinant Pex5p and green fluorescent protein (GFP) carrying a peroxisome-targeting signal were co-injected into the mutant cells, the GFP fluorescence gathered over time to particulate structures where PMP70 was co-localized. This process was dependent on both Pex5p and the targeting signal, and, most importantly, occurred even in the presence of cycloheximide, a protein synthesis inhibitor. These findings suggest that the ghosts act as acceptors of matrix proteins in the peroxisome recovery process at least in the PEX5 mutant, and support the view that peroxisomes can grow by incorporating newly synthesized matrix proteins.

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Biogenesis of Peroxisomal Proteins in Vivo and in Vitro*

Cited by 85 publications

References 13 publications

Peroxisomes in wild-type and rosy mutant Drosophila melanogaster.

Peroxisomes in wild-type and rosy mutant Drosophila melanogaster.

Insertion of the 70-kDa Peroxisomal Membrane Protein into Peroxisomal Membranes in Vivo and in Vitro

Formation of Peroxisomes from Peroxisomal Ghosts in a Peroxisome-deficient Mammalian Cell Mutant upon Complementation by Protein Microinjection

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