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| 著者 | Tuncel A., Kawaguchi J., Ihara Y., Matsusaka H., Nishi A., Nakamura T., Kuhara S., Hirakawa H., Nakamura Y., Cakir B., Nagamine A., Okita T.W., Hwang S.K., Satoh H. | |
| タイトル | The rice endosperm ADP-glucose pyrophosphorylase large subunit is essential for optimal catalysis and allosteric regulation of the heterotetrameric enzyme. | |
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Abstract: Although an alternative pathway has been suggested, the prevailing view is that starch synthesis in cereal endosperm is controlled by the activity of the cytosolic isoform of ADPglucose pyrophosphorylase (AGPase). In rice, the cytosolic AGPase isoform is encoded by the OsAGPS2b and OsAGPL2 genes, which code for the small (S2b) and large (L2) subunits of the heterotetrameric enzyme, respectively. In this study, we isolated several allelic missense and nonsense OsAGPL2 mutants by N-methyl-N-nitrosourea (MNU) treatment of fertilized egg cells and by TILLING (Targeting Induced Local Lesions in Genomes). Interestingly, seeds from three of the missense mutants (two containing T139I and A171V) were severely shriveled and had seed weight and starch content comparable with the shriveled seeds from OsAGPL2 null mutants. Results from kinetic analysis of the purified recombinant enzymes revealed that the catalytic and allosteric regulatory properties of these mutant enzymes were significantly impaired. The missense heterotetramer enzymes and the S2b homotetramer had lower specific (catalytic) activities and affinities for the activator 3-phosphoglycerate (3-PGA). The missense heterotetramer enzymes showed more sensitivity to inhibition by the inhibitor inorganic phosphate (Pi) than the wild-type AGPase, while the S2b homotetramer was profoundly tolerant to Pi inhibition. Thus, our results provide definitive evidence that starch biosynthesis during rice endosperm development is controlled predominantly by the catalytic activity of the cytoplasmic AGPase and its allosteric regulation by the effectors. Moreover, our results show that the L2 subunit is essential for both catalysis and allosteric regulatory properties of the heterotetramer enzyme. |
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| 掲載誌 | Plant Cell Physiol. | |
| Country | unknown | |
| 刊数 | 55(6) | |
| ページ | 1169-83 | |
| 発刊年 | 2014 | |
| PubMed ID | 24747952 | |
| PubMed Central ID | - | |
| DOI | 10.1093/pcp/pcu057 | |
| URL | - | |
| 関連情報 | ||
| 遺伝子 | AGPL2 AGPS2 | |
| INSD | - | |
| 系統 |
野生イネ コアコレクション |
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| 突然変異系統(遺伝研 Collection) | - | |
| 種子不稔系統 | - | |
| 致死変異体系統 | - | |
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組織別発生段階 - 突然変異系統(遺伝子) |
- | |
| 栽培イネ品種(遺伝研 Collection) | - | |
| 組織別発生段階 | - | |
