TY - JOUR A1 - Dauvillee, David A1 - Chochois, Vincent A1 - Steup, Martin A1 - Haebel, Sophie A1 - Eckermann, Nora A1 - Ritte, Gerhard A1 - Ral, Jean-Philippe A1 - Colleoni, Christophe A1 - Hicks, Glenn A1 - Wattebled, Fabrice A1 - Deschamps, Philippe A1 - Lienard, Luc A1 - Cournac, Laurent A1 - Putaux, Jean-Luc A1 - Dupeyre, Danielle A1 - Ball, Steven G. T1 - Plastidial phosphorylase is required for normal starch synthesis in Chlamydomonas reinhardtii JF - The plant journal N2 - Among the three distinct starch phosphorylase activities detected in Chlamydomonas reinhardtii, two distinct plastidial enzymes (PhoA and PhoB) are documented while a single extraplastidial form (PhoC) displays a higher affinity for glycogen as in vascular plants. The two plastidial phosphorylases are shown to function as homodimers containing two 91-kDa (PhoA) subunits and two 110-kDa (PhoB) subunits. Both lack the typical 80-amino-acid insertion found in the higher plant plastidial forms. PhoB is exquisitely sensitive to inhibition by ADP-glucose and has a low affinity for malto-oligosaccharides. PhoA is more similar to the higher plant plastidial phosphorylases: it is moderately sensitive to ADP-glucose inhibition and has a high affinity for unbranched malto-oligosaccharides. Molecular analysis establishes that STA4 encodes PhoB. Chlamydomonas reinhardtii strains carrying mutations at the STA4 locus display a significant decrease in amounts of starch during storage that correlates with the accumulation of abnormally shaped granules containing a modified amylopectin structure and a high amylose content. The wild-type phenotype could be rescued by reintroduction of the cloned wild-type genomic DNA, thereby demonstrating the involvement of phosphorylase in storage starch synthesis. KW - Chlamydomonas KW - starch KW - amylopectin KW - (glycogen) starch phosphorylase Y1 - 2006 U6 - https://doi.org/10.1111/j.1365-313X.2006.02870.x SN - 0960-7412 VL - 48 IS - 2 SP - 274 EP - 285 PB - Blackwell CY - Oxford ER - TY - JOUR A1 - Fettke, Jörg A1 - Chia, Tansy A1 - Eckermann, Nora A1 - Smith, Alison M. A1 - Steup, Martin T1 - A transglucosidase necessary for starch degradation and maltose metabolism in leaves at night acts on cytosolic heteroglycans (SHG) N2 - The recently characterized cytosolic transglucosidase DPE2 (EC 2.4.1.25) is essential for the cytosolic metabolism of maltose, an intermediate on the pathway by which starch is converted to sucrose at night. In in vitro assays, the enzyme utilizes glycogen as a glucosyl acceptor but the in vivo acceptor molecules remained unknown. In this communication we present evidence that DPE2 acts on the recently identified cytosolic water-soluble heteroglycans (SHG) as does the cytosolic phosphorylase (EC 2.4.1.1) isoform. By using in vitro two-step C-14 labeling assays we demonstrate that the two transferases can utilize the same acceptor sites of the SHG. Cytosolic heteroglycans from a DPE2-deficient Arabidopsis mutant were characterized. Compared with the wild type the glucose content of the heteroglycans was increased. Most of the additional glucosyl residues were found in the outer chains of SHG that are released by an endo- alpha-arabinanase (EC 3.2.1.99). Additional starch-related mutants were characterized for further analysis of the increased glucosyl content. Based on these data, the cytosolic metabolism of starch-derived carbohydrates is discussed Y1 - 2006 UR - http://www3.interscience.wiley.com/cgi-bin/issn?DESCRIPTOR=PRINTISSN&VALUE=0960-7412 U6 - https://doi.org/10.1111/j.1365-313X.2006.02732.x SN - 0960-7412 ER -