Regulation of efflux pump expression and drug resistance by the 2 transcription factors

31 Constitutive overexpression of the Mdr1 efflux pump is an important mechanism of acquired 32 drug resistance in the yeast Candida albicans. The zinc cluster transcription factor Mrr1 is a 33 central regulator of MDR1 expression, but other transcription factors have also been 34 implicated in MDR1 regulation. To better understand how MDR1-mediated drug resistance is 35 achieved in this fungal pathogen, we studied the interdependence of Mrr1 and two other 36 MDR1 regulators, Upc2 and Cap1, in the control of MDR1 expression. A mutated, 37 constitutively active Mrr1 could upregulate MDR1 and confer drug resistance in the absence 38 of Upc2 or Cap1. On the other hand, Upc2 containing a gain-of-function mutation only 39 slightly activated the MDR1 promoter, and this activation depended on the presence of a 40 functional MRR1 gene. In contrast, a C-terminally truncated, activated form of Cap1 could 41 upregulate MDR1 in a partially Mrr1-independent fashion. The induction of MDR1 expression 42 by toxic chemicals occurred independently of Upc2, but required the presence of Mrr1 and 43 also partially depended on Cap1. Transcriptional profiling and in vivo DNA binding studies 44 showed that a constitutively active Mrr1 binds to and upregulates most of its direct target 45 genes in the presence or absence of Cap1. Therefore, Mrr1 and Cap1 cooperate in the 46 environmental induction of MDR1 expression in wild-type C. albicans, but gain-of-function 47 mutations in either of the two transcription factors can independently mediate efflux pump 48 overexpression and drug resistance. 49 on A uust 9, 2017 by gest httpaac.asm .rg/ D ow nladed fom