title: Non-coding nucleotides and amino acids near the active site regulate peptide deformylase expression and inhibitor susceptibility in Chlamydia trachomatis.
abstract: Chlamydia trachomatis, an obligate intracellular bacterium, is a highly prevalent human pathogen. Hydroxamic-acid-based matrix metalloprotease inhibitors can effectively inhibit the pathogen both in vitro and in vivo, and have exhibited therapeutic potential. Here, we provide genome sequencing data indicating that peptide deformylase (PDF) is the sole target of the inhibitors in this organism. We further report molecular mechanisms that control chlamydial PDF (cPDF) expression and inhibition efficiency. In particular, we identify the σ⁶⁶-dependent promoter that controls cPDF gene expression and demonstrate that point mutations in this promoter lead to resistance by increasing cPDF transcription. Furthermore, we show that substitution of two amino acids near the active site of the enzyme alters enzyme kinetics and protein stability.
authors: Department of Physiology and Biophysics, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, NJ 08854, USA.
authors: Xiaofeng
authors: X
authors: Bao
authors: Department of Physiology and Biophysics, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, NJ 08854, USA.
authors: Niseema D
authors: ND
authors: Pachikara
authors: Department of Physiology and Biophysics, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, NJ 08854, USA.
authors: Christopher B
authors: CB
authors: Oey
authors: Department of Physiology and Biophysics, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, NJ 08854, USA.
authors: Amit
authors: A
authors: Balakrishnan
authors: Laboratory of Molecular Biophysics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
authors: Lars F
authors: LF
authors: Westblade
authors: Department of Microbiology and Molecular Genetics, and Department of Medicine, University of California, Irvine, CA 92697, USA.
authors: Ming
authors: M
authors: Tan
authors: Department of Biochemistry and Microbiology, School of Environmental and Biological Science, Rutgers, The State University of New Jersey, New Brunswick, NJ 08901, USA.
authors: Theodore
authors: T
authors: Chase
authors: Department of Genetics and Waksman Institute, The State University of New Jersey, Piscataway, NJ 08854, USA.
authors: Bryce E
authors: BE
authors: Nickels
authors: Department of Physiology and Biophysics, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, NJ 08854, USA.
authors: Huizhou
authors: H
authors: Fan
mesh_terms: D024506:5' Flanking Region; D000581:Amidohydrolases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*; D019943:Amino Acid Substitution; D001483:Base Sequence; D020134:Catalytic Domain; D002692:Chlamydia trachomatis / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D004151:Dipeptides / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000187:drug effects / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D009154:Mutation / Q000235:genetics; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
chemical_list: D004151:Dipeptides; D004791:Enzyme Inhibitors; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D000581:Amidohydrolases; C084507:peptide deformylase