title: A genetic model to investigate drug-target interactions at the ribosomal decoding site.
abstract: Recent advances in X-ray crystallography have greatly contributed to the understanding of the structural interactions between aminoglycosides and the ribosomal decoding site. Efforts to genetically probe the functional relevance of proposed drug-nucleotide contacts have in part been hampered by the presence of multiple rRNA operons in most bacteria. A derivative of the Gram-positive Mycobacterium smegmatis was rendered single rRNA operon allelic by means of gene inactivation techniques. In this system, genetic manipulation of the single chromosomal rRNA operon results in cells carrying homogeneous populations of mutant ribosomes. An exhaustive mutagenesis study of the ribosomal A site has been performed to define the importance of individual drug-nucleotide contacts. Mutational alterations in the M. smegmatis decoding site are discussed here, comparing the results with those obtained in other organisms. Implications for the selectivity of antimicrobial agents and for the fitness cost of resistance mutations are addressed.
authors.affiliation: Institut für Medizinische Mikrobiologie, Universität Zürich, Gloriastrasse 32, 8006 Zurich, Switzerland.
authors.forename: S N
authors.initials: SN
authors.lastname: Hobbie
authors.forename: C
authors.initials: C
authors.lastname: Bruell
authors.forename: S
authors.initials: S
authors.lastname: Kalapala
authors.forename: S
authors.initials: S
authors.lastname: Akshay
authors.forename: S
authors.initials: S
authors.lastname: Schmidt
authors.forename: P
authors.initials: P
authors.lastname: Pfister
authors.forename: E C
authors.initials: EC
authors.lastname: Böttger
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites / Q000235:genetics; D008957:Models, Genetic; D008958:Models, Molecular; D015394:Molecular Structure; D009154:Mutation / Q000235:genetics; D012329:RNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S