The common aromatic amino acid biosynthesis pathway is essential in Mycobacterium tuberculosis


Parish, T; Stoker, NG; (2002) The common aromatic amino acid biosynthesis pathway is essential in Mycobacterium tuberculosis. Microbiology (Reading, England), 148 (Pt 10). pp. 3069-77. ISSN 1350-0872

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Abstract

Attempts to construct Mycobacterium tuberculosis strains with a defect in the common aromatic amino acid biosynthesis pathway were made. In other bacteria the genes of this pathway (aro) can be disrupted in the presence of suitable media supplements. The genomic organization of the aro genes in M. tuberculosis reveals that there is one operon (aroCKBQ) and three isolated aro genes (aroE, aroG and aroA). The aroK gene was chosen as a target for disruption; this encodes shikimate kinase, which catalyses the fifth step in chorismate biosynthesis. Attempts to replace the wild-type aroK gene with a disrupted allele (aroKDelta::hyg) by a two-step homologous recombination procedure were unsuccessful in a wild-type strain. When a second functional copy of aroK was integrated into the chromosome, it was possible to isolate a strain carrying the disrupted gene. Excision of the L5-integrated copy of aroK by the L5 excisionase could be not be achieved in the strain carrying the disrupted copy, but was possible in a strain carrying a wild-type copy. These results demonstrate that the chorismate pathway is essential for the viability of M. tuberculosis.

Item Type: Article
Keywords: Amino Acids, Aromatic/biosynthesis, Bacterial Proteins/genetics/metabolism, Chorismic Acid/*metabolism, DNA Nucleotidyltransferases, Gene Deletion, *Genes, Essential, Mycobacteriophages/enzymology, Mycobacterium tuberculosis/genetics/*growth & development/metabolism, Operon, Phosphotransferases (Alcohol Group Acceptor)/*genetics/metabolism, Recombination, Genetic, Support, Non-U.S. Gov't, Transformation, Bacterial
Faculty and Department: Faculty of Infectious and Tropical Diseases > Dept of Pathogen Molecular Biology
PubMed ID: 12368440
Web of Science ID: 178588300017
URI: http://researchonline.lshtm.ac.uk/id/eprint/16514

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