Identification of the cellulose synthase genes from the Oomycete Saprolegnia monoica and effect of cellulose synthesis inhibitors on gene expression and enzyme activity.
Cellulose biosynthesis is a vital but yet poorly understood biochemical process in Oomycetes. Here, we report the identification and characterization of the cellulose synthase genes (CesA) from Saprolegnia monoica. Southern blot experiments revealed the occurrence of three CesA homologues in this species and phylogenetic analyses confirmed that Oomycete CesAs form a clade of their own. All gene products contained the D,D,D,QXXRW signature of most processive glycosyltransferases, including cellulose synthases. However, their N-terminal ends exhibited Oomycete-specific domains, i.e. Pleckstrin Homology domains, or conserved domains of an unknown function together with additional putative transmembrane domains. Mycelial growth was inhibited in the presence of the cellulose biosynthesis inhibitors 2,6-dichlorobenzonitrile or Congo Red. This inhibition was accompanied by a higher expression of all CesA genes in the mycelium and increased in vitro glucan synthase activities. Altogether, our data strongly suggest a direct involvement of the identified CesA genes in cellulose biosynthesis.
Fugelstad
Johanna
J
Division of Glycoscience, School of Biotechnology, Royal Institute of Technology, AlbaNova University Centre, Stockholm, Sweden.
Bouzenzana
Jamel
J
Djerbi
Soraya
S
Guerriero
Gea
G
Ezcurra
Inés
I
Teeri
Tuula T
TT
Arvestad
Lars
L
Bulone
Vincent
V
D020418:Algal Proteins / Q000235:genetics / Q000378:metabolism; D015139:Blotting, Southern; D002482:Cellulose / Q000096:biosynthesis*; D003224:Congo Red / Q000494:pharmacology; D048028:DNA, Algal / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005786:Gene Expression Regulation; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology; D044762:Saprolegnia / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D013312:Stress, Physiological
D020418:Algal Proteins; D048028:DNA, Algal; D004791:Enzyme Inhibitors; D009570:Nitriles; D003224:Congo Red; D002482:Cellulose; D005964:Glucosyltransferases; C478648:cellulose synthase; C055869:dichlobanil