Functional characterization of the upstream components of the Hog1-like kinase cascade in hyperosmotic and carbon...

Trichoderma reesei holds a high capacity for protein secretion and represents the most important cellulase producer in industry. However, the external signal sensing and intracellular signal transduction during cellulose induction remain unclear. As one of the most pervasive signal transduction pathways in all eukaryotic species, the mitogen-activated protein kinase (MAPK) pathway and its upstream sensing and signaling components are involved in various physiological processes including stress and nutrient sensing. Particularly, the Hog1-type MAPK Tmk3 has been reported to be involved in the cellulase production in T. reesei.

Here we established the physiological role of two upstream regulatory branches, the Sho1 branch and the Sln1 branch, of the Hog1-type Tmk3 pathway in T. reesei. Deletion of Trste20 of the Sho1 branch or repression of Trypd1 of the Sln1 branch reduced the resistance to high salt stress, whereas TrSho1 showed an opposing effect to that of TrSte20 and the identified TrSln1 seemed to be dispensable in the osmotic regulation. The Sho1 and Sln1 branches also participated in the cell wall integrity maintenance and other stress responses (i.e. oxidative and thermo stresses). Notably, TrSho1 and TrSte20 of the Sho1 branch and TrYpd1 of the Sln1 branch were shown to be differentially involved in the cellulase production of T. reesei. Repression of Trypd1 hardly affected cellulase induction, whereas overexpression of Trypd1 resulted in the reduced production of cellulases. Contrary to the case of Trypd1, repression of Trsho1 or deletion of Trste20 significantly reduced the transcription of cellulase genes.

TrSho1 and TrSte20 of the Sho1 branch and TrYpd1 of the Sln1 branch are all involved in general stress responses including hyperosmotic regulation and cell wall integrity maintenance. Moreover, our study revealed that the Sho1 and Sln1 osmosensing pathways are differentially involved in the regulation of cellulase production in T. reesei. The Sho1 branch positively regulated the production of cellulases and the transcription of cellulase genes while TrYpd1 of the Sln1 branch negatively controlled the cellulase production, supporting the crosstalks of osmosensing and nutrient sensing.