Scm3 is essential for stability of CENPA in C. albicans. (A) The C. albicans strain CaAB2 (SCM3/scm3::FRT), where one copy of SCM3 has been deleted, and two independent transformants CaAB3 and CaAB4 (MET3prSCM3/scm3::FRT), where the remaining allele was placed under the control of the MET3 promoter, were streaked on plates containing permissive (CM-met-cys) and nonpermissive (CM + 1 mM met + 1 mM cys) media and photographed after 48 h of incubation at 30°C. (B) Western blot showing protein levels of CENPA upon depletion of Scm3 for the indicated time points normalized to PSTAIRE. (C) Dissociation of the CENPA (GFP) cluster marking disintegration of the kinetochore in CaAB7 (MET3prSCM3/scm3::FRT CSE4-GFP-CSE4/CSE4) cells grown in nonpermissive conditions for the indicated time. Scale bar, 5 µm. n = 100. (D) ChIP-qPCR using anti-GFP (CENPA) antibodies revealed reduced CENPA enrichment at CEN7 in CaAB7 when grown in nonpermissive media for 8 h. Two-way ANOVA; (***) P-value < 0.001; (ns) P-value > 0.05; n = 3.
