Toward an understanding of the biogenesis of the starch granule. Determination of granule-bound and soluble starch synthase functions in amylopectin synthesis.
Plant starch synthesis can be distinguished from those of bacterial, fungal, and animal glycogen by the presence of multiple elongation (starch synthases) and branching enzymes. This complexity has precluded genetic assignment of functions to the various soluble starch synthases in the building of amylopectin. In Chlamydomonas, we have recently shown that defects in the major soluble starch synthase lead to a specific decrease in the amount of a subset of amylopectin chains whose length ranges between 8 and 40 glucose residues (Fontaine, T., D'Hulst, C., Maddelein, M.-L., Routier, F., Marianne-Pepin, T., Decq, A., Wieruszeski, J. M., Delrue, B., Van Den Koornhuyse, N., Bossu, J.-P., Fournet, B., and Ball, S. G. (1993) J. Biol. Chem. 268, 16223-16230). We now demonstrate that granule-bound starch synthase, the enzyme that was thought to be solely responsible for amylose synthesis, is involved in amylopectin synthesis. Disruption of the Chlamydomonas granule-bound starch synthase structural gene establishes that synthesis of long chains by this enzyme can become an absolute requirement for amylopectin synthesis in particular mutant backgrounds. In the sole presence of soluble starch synthase I, Chlamydomonas directs the synthesis of a major water-soluble polysaccharide fraction and minute amounts of a new type of highly branched granular material, whose structure is intermediate between those of glycogen and amylopectin. These results lead us to propose that the nature of the elongation enzyme conditions the synthesis of distinct size classes of glucans in all starch fractions.
Maddelein
M L
ML
Laboratoire de Chimie Biologique, Unité Mixte de Recherche du CNRS 111, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Libessart
N
N
Bellanger
F
F
Delrue
B
B
D'Hulst
C
C
Van den Koornhuyse
N
N
Fontaine
T
T
Wieruszeski
J M
JM
Decq
A
A
Ball
S
S
D000687:Amylopectin / Q000096:biosynthesis*; D000818:Animals; D002696:Chlamydomonas / Q000201:enzymology* / Q000235:genetics; D009154:Mutation; D011134:Polysaccharides / Q000737:chemistry; D013213:Starch / Q000096:biosynthesis*; D013214:Starch Synthase / Q000235:genetics / Q000502:physiology*
D011134:Polysaccharides; D013213:Starch; D000687:Amylopectin; D013214:Starch Synthase