RRC ID 33338
著者 Seung D, Soyk S, Coiro M, Maier BA, Eicke S, Zeeman SC.
タイトル PROTEIN TARGETING TO STARCH is required for localising GRANULE-BOUND STARCH SYNTHASE to starch granules and for normal amylose synthesis in Arabidopsis.
ジャーナル PLoS Biol
Abstract The domestication of starch crops underpinned the development of human civilisation, yet we still do not fully understand how plants make starch. Starch is composed of glucose polymers that are branched (amylopectin) or linear (amylose). The amount of amylose strongly influences the physico-chemical behaviour of starchy foods during cooking and of starch mixtures in non-food manufacturing processes. The GRANULE-BOUND STARCH SYNTHASE (GBSS) is the glucosyltransferase specifically responsible for elongating amylose polymers and was the only protein known to be required for its biosynthesis. Here, we demonstrate that PROTEIN TARGETING TO STARCH (PTST) is also specifically required for amylose synthesis in Arabidopsis. PTST is a plastidial protein possessing an N-terminal coiled coil domain and a C-terminal carbohydrate binding module (CBM). We discovered that Arabidopsis ptst mutants synthesise amylose-free starch and are phenotypically similar to mutants lacking GBSS. Analysis of granule-bound proteins showed a dramatic reduction of GBSS protein in ptst mutant starch granules. Pull-down assays with recombinant proteins in vitro, as well as immunoprecipitation assays in planta, revealed that GBSS physically interacts with PTST via a coiled coil. Furthermore, we show that the CBM domain of PTST, which mediates its interaction with starch granules, is also required for correct GBSS localisation. Fluorescently tagged Arabidopsis GBSS, expressed either in tobacco or Arabidopsis leaves, required the presence of Arabidopsis PTST to localise to starch granules. Mutation of the CBM of PTST caused GBSS to remain in the plastid stroma. PTST fulfils a previously unknown function in targeting GBSS to starch. This sheds new light on the importance of targeting biosynthetic enzymes to sub-cellular sites where their action is required. Importantly, PTST represents a promising new gene target for the biotechnological modification of starch composition, as it is exclusively involved in amylose synthesis.
巻・号 13(2)
ページ e1002080
公開日 2015-2-1
DOI 10.1371/journal.pbio.1002080
PII PBIOLOGY-D-14-02490
PMID 25710501
PMC PMC4339375
MeSH Amylopectin / metabolism Amylose / biosynthesis* Arabidopsis / classification Arabidopsis / genetics* Arabidopsis / metabolism Arabidopsis Proteins / genetics* Arabidopsis Proteins / metabolism Breeding Cytoplasmic Granules / chemistry Cytoplasmic Granules / metabolism Gene Expression Regulation, Plant* Phylogeny Plant Leaves / genetics Plant Leaves / metabolism Plants, Genetically Modified Protein Structure, Tertiary Starch Synthase / genetics* Starch Synthase / metabolism
IF 7.076
引用数 59
WOS 分野 BIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
シロイヌナズナ / 植物培養細胞・遺伝子 pda04554