title: Deletion of cyp125 Confers Increased Sensitivity to Azoles in Mycobacterium tuberculosis.
abstract: Mycobacterium tuberculosis is able to utilize cholesterol as a carbon source, and this ability is linked to its virulence in macrophages and in the mouse model of infection. The M. tuberculosis cytochrome P450 Cyp125 plays a key role in cholesterol metabolism being involved in the first steps of its degradation. Cyp125 is a cholesterol hydroxylase which is essential for cholesterol catabolism in M. bovis BCG and some strains of M. tuberculosis. We generated an unmarked, in-frame deletion of Cyp125 in M. tuberculosis H37Rv. The deletion strain was able to grow as well as wild-type in medium containing glucose as the carbon source. The Cyp125 deletion strain was more sensitive to growth inhibition by clotrimazole consistent with the ability of Cyp125 to bind azoles with high affinity. The deletion strain showed no difference in sensitivity to nitric oxide or hydrogen peroxide and was not attenuated for growth inside THP-1 human macrophage-like cells. These data suggest that the attenuation of virulence seen in operon deletion strains is not linked to the lack of Cyp125 alone.
authors.affiliation: Queen Mary University of London, Barts & The London School of Medicine and Dentistry, Centre for Infectious Disease, London, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Carroll
authors.affiliation: Queen Mary University of London, Barts & The London School of Medicine and Dentistry, Centre for Infectious Disease, London, United Kingdom; TB Discovery Research, Infectious Disease Research Institute, Seattle, Washington, United States of America.
authors.forename: Tanya
authors.initials: T
authors.lastname: Parish
mesh_terms: D001393:Azoles / Q000737:chemistry*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002244:Carbon / Q000737:chemistry; D002460:Cell Line; D002784:Cholesterol / Q000737:chemistry; D003022:Clotrimazole / Q000737:chemistry; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D017353:Gene Deletion; D005947:Glucose / Q000737:chemistry; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology*; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics*; D009569:Nitric Oxide / Q000737:chemistry; D014774:Virulence
chemical_list: D001393:Azoles; D001426:Bacterial Proteins; D009569:Nitric Oxide; D002244:Carbon; D003577:Cytochrome P-450 Enzyme System; D002784:Cholesterol; D003022:Clotrimazole; D005947:Glucose