(A) Alignment of C. neoformans CnCna1 and the three calcineurin A catalytic subunit proteins (McCnaA, McCnaB and McCnaC) in M. circinelloides with A. fumigatus CnaA SPRR (AfCnaA). (B) Complementation of the A. fumigatus ΔcnaA mutant with native AfCnaA, C. neoformans CnCNA1, a CNAFCNA chimera consisting of the N-terminus catalytic domain of C. neoformans CNA1 and the C-terminal portion of AfcnaA consisting of the CnBBH, SPRR, CaMBD and the AID, M. circinelloides McCnaA, and MccnaC. (C) Complementation of the A. fumigatus ΔcnaA mutant with M. grisea MgcnaA and N. crassa NccnaA. Strains were assessed for growth complementation and CnaA septal localization (arrow) by fluorescence microscopy. (D) Western detection of the calcineurin A proteins expressed in the complemented strains. (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.