The Mnn2 Mannosyltransferase Family Modulates Mannoprotein Fibril Length, Immune Recognition and Virulence of Candida albicans
Figure 2
The MNN2 gene family is required for cell wall integrity.
(A) Stationary phase cultures were diluted to OD600 0.5 in YPD and 3 µl from 1∶10 serial dilutions spotted onto YPD agar plates containing either 100 µg/ml CFW, 0.03% SDS, 100 µg/ml Congo Red (CR) and incubated at 30°C for 48 h, or spotted on YPD agar and incubated at 42°C. (B) Activation of the cell wall salvage pathway was determined by western blot on crude protein extracts prepared from exponentially growing cells. The PhosphoPlus P44/42 antibody detects phosphorylated (activated) Mkc1. As a positive control for Mkc1 activation wild type (CAI-4+CIp10) cells were exposed to 0.0032 µg/ml caspofungin for 10 min. Lane 1 wild type +CIp10, lane 2 wild type+caspofungin, lane 3 mnn2Δ, lane 4 mnn2Δ+MNN2, lane 5 mnn26Δ, lane 6 mnn26Δ+MNN26, lane 7 mnn2Δ/mnn26Δ, lane 8 mnn2Δ/mnn26Δ+MNN2/MNN26, lane 9 mnn23Δ/mnn26Δ, lane 10 mnn24Δ/mnn26Δ, lane 11 triple mutant, lane 12 quintuple mutant, lane 13 sextuple mutant, lane 14 sextuple mutant+MNN2/MNN26.