Figure 8. Phosphorylation of CnaA is required for proper hyphal growth.

(A) Radial growth after 5 days at 37°C on GMM agar and GMM supplemented with 1.2 M sorbitol (SMM) to assess growth remediation. Wild-type, cnaA^mt-4SA, and cnaA^mt-4SE strains (1×10⁴ conidia each) were cultured on GMM in varying concentrations of caspofungin for 5 days. The cnaA^mt-4SA strain, defective in phosphorylation, exhibits a growth defect and lacks caspofungin-mediated paradoxical growth due to lack of calcineurin activity. (B) Radial growth is depicted as mean colony diameter in triplicate during 1–5 days growth period. (C) Localization of the CnaA mutated constructs by fluorescence microscopy after 24 h growth. (D) Growth reduction of the cnaA^mt-4SA in comparison to the wild-type strain after inoculating 1×10⁶ conidia into GMM liquid and growth for 48 h at 37°C. (E) Calcineurin activity after 24 h growth using p-nitrophenyl phosphate as a substrate. Data from two separate experiments, each with 6 technical replicates are presented as mean ± SD of nanomoles of pNPP released/min/mg protein.