Subcellular localization of five singular WSC domain-containing proteins and their roles in Beauveria bassiana responses to stress cues and metal ions.

Some model fungi have three or four proteins with each vectoring a single cell Wall Stress-responsive Component (WSC) domain at N-terminus. In this study, five proteins, each vectoring only a single WSC domain in N-terminal, central or even C-terminal region, were found in Beauveria bassiana, a filamentous fungal entomopathogen, and named Wsc1A-1E due to the domain singularity. Four of them lack either transmembrane domain or C-terminal conserved signature sequence (DXXD) compared with the homologues in the model fungi. Intriguingly, all the eGFP-tagged fusion proteins of Wsc1A-1E were evidently localized to the cell wall and membrane of transgenic hyphae. Single deletions of the five wsc genes resulted in significant, but differential, increases in cellular sensitivity to cell wall perturbation, oxidation, high osmolarity, and four to six metal ions (Zn(2+) , Mg(2+) , Fe(2+) , K(+) , Ca(2+) and Mn(2+) ). Each deletion mutant also showed a delay of germination and a decrease of conidial UV-B resistance, thermotolerance or both. However, none of the deletions affected substantially the fungal growth, conidiation and virulence. Our results indicate a significance of each WSC protein for the B. bassiana adaptation to diverse habitats of host insects.