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Complementation of a Saccharomyces cerevisiae ERG11/CYP51 (sterol 14α-demethylase) doxycycline-regulated mutant and screening of the azole sensitivity of Aspergillus fumigatus isoenzymes CYP51A and CYP51B. / CM Martel; JE Parker; Warrilow AGS; N Rolley; SL Kelly; DE Kelly; Steven Kelly

Antimicrobial Agents and Chemotherapy, Volume: 54, Issue: 11, Pages: 4920 - 3

Swansea University Author: Steven, Kelly

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DOI (Published version): 10.1128/AAC.00349-10

Abstract

Aspergillus fumigatus sterol 14 alpha-demethylase isoenzymes CYP51A and CYP51B were heterologously expressed in a Saccharomyces cerevisiae mutant (YUG37-erg11), wherein native ERG11/CYP51 expression is controlled using a doxycycline-regulatable promoter. When cultured in the presence of doxycycline,...

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Published in: Antimicrobial Agents and Chemotherapy
Published: American Society of Microbiology 2010
URI: https://cronfa.swan.ac.uk/Record/cronfa9903
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Abstract: Aspergillus fumigatus sterol 14 alpha-demethylase isoenzymes CYP51A and CYP51B were heterologously expressed in a Saccharomyces cerevisiae mutant (YUG37-erg11), wherein native ERG11/CYP51 expression is controlled using a doxycycline-regulatable promoter. When cultured in the presence of doxycycline, recombinant YUG37-pcyp51A and YUG37-pcyp51B yeasts were able to synthesize ergosterol and grow; a control strain harboring reverse-oriented cyp51A could not. YUG37-pcyp51A and YUG37-pcyp51B constructs showed identical sensitivity to itraconazole, posaconazole, clotrimazole, and voriconazole. Conversely, YUG37-pcyp51A withstood 16-fold-higher concentrations of fluconazole than YUG37-pcyp51B (8 and 0.5 mu g ml(-1), respectively).
Keywords: INVASIVE FUNGAL-INFECTIONS; AMINO-ACID SUBSTITUTION; CANDIDA-ALBICANS; ITRACONAZOLE RESISTANCE; FLUCONAZOLE RESISTANCE; REDUCED AFFINITY; AMPHOTERICIN-B; DEMETHYLASE; POSACONAZOLE; ANTIFUNGALS
College: Swansea University Medical School
Issue: 11
Start Page: 4920
End Page: 3