論文 - 詳細
| RRC ID | 11656 |
|---|---|
| 著者 | Funakoshi Y, Negishi Y, Gergen JP, Seino J, Ishii K, Lennarz WJ, Matsuo I, Ito Y, Taniguchi N, Suzuki T. |
| タイトル | Evidence for an essential deglycosylation-independent activity of PNGase in Drosophila melanogaster. |
| ジャーナル | PLoS One |
| Abstract |
BACKGROUND:Peptide:N-glycanase (PNGase) is an enzyme which releases N-linked glycans from glycopeptides/glycoproteins. This enzyme plays a role in the ER-associated degradation (ERAD) pathway in yeast and mice, but the biological importance of this activity remains unknown. PRINCIPAL FINDINGS:In this study, we characterized the ortholog of cytoplasmic PNGases, PNGase-like (Pngl), in Drosophila melanogaster. Pngl was found to have a molecular weight of approximately 74K and was mainly localized in the cytosol. Pngl lacks a CXXC motif that is critical for enzymatic activity in other species and accordingly did not appear to possess PNGase activity, though it still retains carbohydrate-binding activity. We generated microdeletions in the Pngl locus in order to investigate the functional importance of this protein in vivo. Elimination of Pngl led to a serious developmental delay or arrest during the larval and pupal stages, and surviving mutant adult males and females were frequently sterile. Most importantly, these phenotypes were rescued by ubiquitous expression of Pngl, clearly indicating that those phenotypic consequences were indeed due to the lack of functional Pngl. Interestingly, a putative "catalytic-inactive" mutant could not rescue the growth-delay phenotype, indicating that a biochemical activity of this protein is important for its biological function. CONCLUSION:Pngl was shown to be inevitable for the proper developmental transition and the biochemical properties other than deglycosylation activity is important for its biological function. |
| 巻・号 | 5(5) |
| ページ | e10545 |
| 公開日 | 2010-5-10 |
| DOI | 10.1371/journal.pone.0010545 |
| PMID | 20479940 |
| PMC | PMC2866665 |
| MeSH | Amino Acid Sequence Animals Carbohydrate Metabolism Cytosol / enzymology Drosophila melanogaster / enzymology* Drosophila melanogaster / genetics Gene Deletion Gene Expression Regulation, Developmental Glycosylation Homozygote Molecular Sequence Data Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / chemistry Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / genetics Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / metabolism* Phenotype Saccharomyces cerevisiae / enzymology Sequence Homology, Amino Acid Transgenes / genetics |
| IF | 2.74 |
| 引用数 | 26 |
| WOS 分野 | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| ショウジョウバエ | CyO-GFP balancer vg-Gal4 tub-Gal4 and GMR-Gal4 DGRC#109616 |