Ncb2 Is Involved in Activated Transcription of CDR1 in Azole-Resistant Clinical Isolates of Candida albicans

We recently demonstrated thatCDR1 overexpression in azole-resistant isolates ofCandida albicans is due to its enhanced transcriptional activation and increased mRNA stability. In this study, we provide the first evidence of transcriptional regulation ofCDR1 by Ncb2, the β subunit of NC2, a heterodimeric regulator of transcription. ConditionalNCB2 null mutants displayed decreased susceptibility toward azole and an enhanced transcription ofCDR1 . Interestingly, Ncb2 associated with theCDR1 promoter under both repression and activation; however, an increase in recruitment was observed under both transient and constitutive activation states. By chromatin immunoprecipitation (ChIP) assay, we showed the preferential recruitment of Ncb2 to the core TATA region under activation (azole-resistant isolate), while under repression (azole-susceptible isolate) it was present at the TATA upstream region. Further, ChIP analysis revealed that Ncb2 binding was not restricted to theCDR1 gene; instead, it was observed on the promoters of genes coregulated withCDR1 by the transcription activator Tac1. Thetac1 Δ null mutants, which fail to show the drug-induced transient activation ofCDR1 , also showed no increase in Ncb2 recruitment at the promoter. Taken together, our results show that Ncb2, in conjunction with Tac1, is involved in the transcriptional activation ofCDR1 , opening up new therapeutic possibilities to combat multidrug resistance (MDR) inC. albicans .