Selection of transposon mutants of Mycobacterium tuberculosis with increased macrophage infectivity identifies fadD23 to be involved in sulfolipid production and association with macrophages.
Alterations to the composition or architecture of the mycobacterial cell envelope can affect the macrophage infectivity of the bacillus. To further characterize the mycobacterial gene products that modulate the interaction with host cells, we employed transposon mutagenesis and screened for mutants that demonstrated an enhanced binding affinity toward macrophages. After successive rounds of mutant selection and enrichment, a total of five mutants were isolated that harboured gene disruptions within loci involved in lipid synthetic pathways as well as genes coding for putative hypothetical proteins. One mutant in particular, with a disruption in the Rv3826 gene (fadD23), was repeatedly isolated during library screening. Analysis of the cell envelope constituents of the Tn : : fadD23 strain revealed a lack of sulfolipid production which was restored following complementation with the wild-type gene.
Lynett
Jennifer
J
Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V5Z 4H4, Canada.
Stokes
Richard W
RW
Departments of Microbiology and Immunology and Paediatrics, University of British Columbia; Division of Infectious and Immunological Diseases, British Columbia's Children's Hospital, Vancouver, BC V5Z 4H4, Canada.
D002460:Cell Line; D004251:DNA Transposable Elements; D006801:Humans; D008055:Lipids / Q000096:biosynthesis*; D008264:Macrophages / Q000382:microbiology*; D016254:Mutagenesis, Insertional; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*
D004251:DNA Transposable Elements; D008055:Lipids; C015518:sulfolipids