Type‐II 3‐oxoacyl‐CoA thiolase of the nematode <i>Caenorhabditis elegans</i> is located in peroxisomes, highly expressed during larval stages and induced by clofibrate

Maebuchi, Motohiro; Togo, Summnanuna H.; Yokota, Sadaki; Ghenea, Simona; Bun-ya, Masanori; Kamiryo, Tatsuyuki; Kawahara, Akira

doi:10.1046/j.1432-1327.1999.00655.x

Cited by 11 publications

(20 citation statements)

References 34 publications

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“…Afterwards, little attention was paid to nematode peroxisomes. In previous studies, we have shown that type II 3-oxoacyl-CoA thiolase and catalase-2 are immunocytochemically localized to peroxisomes of Caenorhabditis elegans (Maebuchi et al 1999;Togo et al 2000), confirming that peroxisomes exist in the nematode. Caenorhabditis elegans is one of the most important experimental animals in genetic and molecular biology and a study on targeting signals of enzymes in nematode peroxisomes has been started (de Vet and van den Bosch 2000;Gurvitz et al 2001;Motley et al 2002).…”

Section: Introductionsupporting

confidence: 57%

Peroxisomes of the nematode Caenorhabditis elegans: distribution and morphological characteristics

Yokota

Togo

Maebuchi

et al. 2002

Histochem Cell Biol

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We analyzed the distribution and morphological characteristics of peroxisomes in the nematode Caenorhabditis elegans by routine electron microscopy, immunoelectron microscopy, and morphometry. Peroxisomes were mainly contained in the epithelial cells of the digestive tract and pharyngeal gland, but some were observed in other cells. Their shape varied from round to twisted. The matrix of most peroxisomes was coarse and uneven, and contained electron-dense nucleoids and frequently tubular substructures. The diameter of peroxisomes in the gut (0.185 micro m) was smaller than that in pharyngeal gland (0.262 micro m). The volume density of peroxisomes per 100 micro m(2) of cytoplasm was 1.86 in the gut and 1.75 in the pharyngeal gland. After treatment with clofibrate, the diameter of peroxisomes increased approximately 1.11-fold in the gut and 1.2-fold in the pharyngeal gland. The volume density of peroxisomes also increased by 2.2-fold in the gut and 2.6-fold in the pharyngeal gland. The labeling density for catalase-2 was almost identical between gut and pharyngeal gland peroxisomes. The results show that in C. elegans peroxisomes mainly distribute in the epithelial cells of the gut and pharyngeal gland. Peroxisomes of the pharyngeal gland are larger than those of the gut, but peroxisomes of both tissues contain catalase-2 at similar concentrations.

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Section: Introductionsupporting

confidence: 57%

Peroxisomes of the nematode Caenorhabditis elegans: distribution and morphological characteristics

Yokota

Togo

Maebuchi

et al. 2002

Histochem Cell Biol

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“…Although conditions for DAB staining of C. elegans peroxisomes at a lower pH have not yet been established, they were detectable with anti‐(catalase‐2) serum. Using this method, type‐II thiolase of this organism was demonstrated to be a peroxisomal enzyme [22] similar to its mammalian counterparts.…”

Section: Discussionmentioning

confidence: 99%

“…[22]. Procedures for electron microscopy, immunostaining with primary antibodies and secondary gold probes, and determination of the labeling density (gold particles per µm 2 ) were as described previously [22]. Sequences similar to catalase‐2 were searched and analyzed using the ssearch program of DDBJ.…”

Section: Methodsmentioning

confidence: 99%

Immunological detection of alkaline‐diaminobenzidine‐negativeperoxisomes of the nematode Caenorhabditis elegans

Togo

Maebuchi

Yokota³

et al. 2000

European Journal of Biochemistry

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We purified catalase-2 of the nematode Caenorhabditis elegans and identified peroxisomes in this organism. The peroxisomes of C. elegans were not detectable by cytochemical staining using 3, 3'-diaminobenzidine, a commonly used method depending on the peroxidase activity of peroxisomal catalase at pH 9 in which genuine peroxidases are inactive. The cDNA sequences of C. elegans predict two catalases very similar to each other throughout the molecule, except for the short C-terminal sequence; catalase-2 (500 residues long) carries a peroxisomal targeting signal 1-like sequence (Ser-His-Ile), whereas catalase-1 does not. The catalase purified to near homogeneity from the homogenate of C. elegans cells consisted of a subunit of 57 kDa and was specifically recognized by anti-(catalase-2) serum but not by anti-(catalase-1) serum. Subcellular fractionation and indirect immunoelectron microscopy of the nematode detected catalase-2 inside vesicles judged to be peroxisomes using morphological criteria. The purified enzyme (220 kDa) was tetrameric, similar to many catalases from various sources, but exhibited unique pH optima for catalase (pH 6) and peroxidase (pH 4) activities; the latter value is unusually low and explains why the peroxidase activity was undetectable using the standard alkaline diaminobenzidine-staining method. These results indicate that catalase-2 is peroxisomal and verify that it can be used as a marker enzyme for C. elegans peroxisomes.

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“…Finally, the results using the nematode PEX-TPR domain revealed that SKI did not behave as a negative interacting partner compared with SKL (bottom cluster; Table 4). This was not very surprising because C. elegans contains at least one SKI-terminated peroxisomal protein, P-44, that is thought to be involved in phytanic acid degradation and bile acid synthesis (Bun-Ya et al 1997;Maebuchi et al 1999). Both HKL and HRL were at least as ecient as SKL in interacting with the nematode PEX-5 TPR domain (3.0-and 1.8-fold more ecient than SKI).…”

Section: Resultsmentioning

confidence: 99%

The tetratricopeptide repeat domains of human, tobacco, and nematode PEX5 proteins are functionally interchangeable with the analogous native domain for peroxisomal import of PTS1-terminated proteins in yeast

et al. 2001

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In the yeast Saccharomyces cerevisiae, beta-oxidation of fatty acids is compartmentalised in peroxisomes. Most yeast peroxisomal matrix proteins contain a type 1C-terminal peroxisomal targeting signal (PTS1) consisting of the tripeptide SKL or a conservative variant thereof. PTS1-terminated proteins are imported by Pex5p, which interacts with the targeting signal via a tetratricopeptide repeat (TPR) domain. Yeast cells devoid of Pex5p are unable to import PTS1-containing proteins and cannot degrade fatty acids. Here, the PEX5-TPR domains from human, tobacco, and nematode were inserted into a TPR-less yeast Pex5p construct to generate Pex5p chimaeras. These hybrid proteins were examined for functional complementation of the pex5delta mutant phenotype. Expression of the Pex5p chimaeras in pex5delta mutant cells restored peroxisomal import of PTS1-terminated proteins. Chimaera expression also re-established degradation of oleic acid, allowing growth on this fatty acid as a sole carbon source. We conclude that, in the context of Pex5p chimaeras, the human, tobacco, and nematode Pex5p-TPR domains are functionally interchangeable with the native domain for the peroxisomal import of yeast proteins terminating with canonical PTS1s. Non-conserved yeast PTS1s, such as HRL and HKL, did not interact with the tobacco PEX5-TPR domain in the two-hybrid system. HRL occurs at the C-terminus of the peroxisomal protein Eci1p, which is required for growth on unsaturated fatty acids. Although mutant pex5delta cells expressing a yeast/tobacco Pex5p chimaera failed to import a GFP-Eci1p reporter protein, they were able to grow on oleic acid. We reason that this is due to a cryptic PTS in native Eci1p that can function in a redundant system with the C-terminal HRL.

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Type‐II 3‐oxoacyl‐CoA thiolase of the nematode Caenorhabditis elegans is located in peroxisomes, highly expressed during larval stages and induced by clofibrate

Cited by 11 publications

References 34 publications

Peroxisomes of the nematode Caenorhabditis elegans: distribution and morphological characteristics

Peroxisomes of the nematode Caenorhabditis elegans: distribution and morphological characteristics

Immunological detection of alkaline‐diaminobenzidine‐negativeperoxisomes of the nematode Caenorhabditis elegans

The tetratricopeptide repeat domains of human, tobacco, and nematode PEX5 proteins are functionally interchangeable with the analogous native domain for peroxisomal import of PTS1-terminated proteins in yeast

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