Cellulase induction in Trichoderma reesei by cellulose requires its own basal expression.
The induction of cellulases by cellulose, an insoluble polymer, in the filamentous fungus Trichoderma reesei is puzzling. We previously proposed a mechanism that is based on the presence of low levels of cellulase in the uninduced fungus; this basal cellulase activity would digest cellulose-releasing oligosaccharides that could enter the cell and trigger expression of cellulases. We now present experiments that lend further support to this model. We show here that transcripts of two members of the cellulase system, cbh1 and egl1, are present in uninduced T. reesei cells. These transcripts are induced at least 1100-fold in the presence of cellulose. We also show that a construct containing the hygromycin B resistance-encoding gene driven by the cbh1 promoter confers hygromycin B resistance to T. reesei cells grown in the absence of cellulose. Moreover, cellulose-induced production of the cbh1 transcript was suppressed when antisense RNA against three members of the cellulase system was expressed in vivo. Experiments are presented indicating that extracellular cellulase activity is the rate-limiting event in induction of synthesis of the cellulase transcripts by cellulose. The results reveal a critical requirement for basal expression of the cellulase system for induction of synthesis of its own transcripts by cellulose.
Carle-Urioste
J C
JC
Department of Biochemistry, Institute of Chemistry, University of São Paulo, Av. Prof. Lineu Prestes 748, São Paulo 05508-900, SP, Brazil.
Escobar-Vera
J
J
El-Gogary
S
S
Henrique-Silva
F
F
Torigoi
E
E
Crivellaro
O
O
Herrera-Estrella
A
A
El-Dorry
H
H
D001345:Autoradiography; D001483:Base Sequence; D015152:Blotting, Northern; D002480:Cellulase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism*; D043366:Cellulose 1,4-beta-Cellobiosidase; D018076:DNA, Complementary / Q000737:chemistry; D004790:Enzyme Induction; D015971:Gene Expression Regulation, Enzymologic; D005936:Glucans / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D016372:RNA, Antisense / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014242:Trichoderma / Q000201:enzymology*
D018076:DNA, Complementary; D005936:Glucans; D016372:RNA, Antisense; D012333:RNA, Messenger; D002482:Cellulose; D002480:Cellulase; C090774:endoglucanase EGL1; D043366:Cellulose 1,4-beta-Cellobiosidase; C003328:sophorose