Starchless mutants of Chlamydomonas reinhardtii lack the small subunit of a heterotetrameric ADP-glucose pyrophosphorylase.
ADP-glucose synthesis through ADP-glucose pyrophosphorylase defines the major rate-controlling step of storage polysaccharide synthesis in both bacteria and plants. We have isolated mutant strains defective in the STA6 locus of the monocellular green alga Chlamydomonas reinhardtii that fail to accumulate starch and lack ADP-glucose pyrophosphorylase activity. We show that this locus encodes a 514-amino-acid polypeptide corresponding to a mature 50-kDa protein with homology to vascular plant ADP-glucose pyrophosphorylase small-subunit sequences. This gene segregates independently from the previously characterized STA1 locus that encodes the large 53-kDa subunit of the same heterotetramer enzyme. Because STA1 locus mutants have retained an AGPase but exhibit lower sensitivity to 3-phosphoglyceric acid activation, we suggest that the small and large subunits of the enzyme define, respectively, the catalytic and regulatory subunits of AGPase in unicellular green algae. We provide preliminary evidence that both the small-subunit mRNA abundance and enzyme activity, and therefore also starch metabolism, may be controlled by the circadian clock.
Zabawinski
C
C
Laboratoire de Chimie Biologique, Unité Mixte de Recherche du C.N.R.S. No. 8576, Université des Sciences et Technologies de Lille, 59655 Villeneuve d'Ascq Cedex, France.
Van Den Koornhuyse
N
N
D'Hulst
C
C
Schlichting
R
R
Giersch
C
C
Delrue
B
B
Lacroix
J M
JM
Preiss
J
J
Ball
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001683:Biological Clocks; D002459:Cell Fusion; D016825:Chlamydomonas reinhardtii / Q000201:enzymology / Q000235:genetics*; D002940:Circadian Rhythm; D003001:Cloning, Molecular; D051599:Glucose-1-Phosphate Adenylyltransferase; D008969:Molecular Sequence Data; D009154:Mutation; D009713:Nucleotidyltransferases / Q000145:classification / Q000235:genetics*; D010802:Phylogeny; D011123:Polyploidy; D020836:Protein Structure, Quaternary; D011523:Protoplasts; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D012641:Selection, Genetic; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013213:Starch / Q000096:biosynthesis*
D012333:RNA, Messenger; D018749:RNA, Plant; D013213:Starch; D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase