The lpqS knockout mutant of Mycobacterium tuberculosis is attenuated in Macrophages

Lipoproteins of Mycobacterium tuberculosis (M. tuberculosis) represent an important class of cell envelop proteins. On the whole, 99 putative lipoproteins have been identified in the genome of M. tuberculosis. Earlier investigations on individual mycobacterial lipoproteins demonstrate that some lipoproteins elicit a strong immune response, while others are virulence factors in different models of infection. LpqS is an uncharacterized lipoprotein encoded by the open reading frame Rv0847 of M. tuberculosis. In the present study, we have characterized this putative lipoprotein LpqS with respect to the virulence of M. tuberculosis. A mutant of the M. tuberculosis H(37)Rv strain not producing LpqS (Delta lpqS) was generated by specialized transduction. The deletion mutant showed reduced growth in Sauton's minimal media and was highly sensitive to SOS and copper, compared to the wild type when grown on solid media. In vitro infection studies showed that the mutant was attenuated for growth in PMA-activated THP-1 cells. Complementation of the mutant with a single copy of the gene cloned under the hsp60 promoter partially restored the phenotype of the wild type strain H(37)Rv. Thus INS plays an important role in sensing the host macrophage environment and might be required for the intracellular survival of M. tuberculosis. Cotranscription of INS with the genes downstream cysK2, Rv0849 and Rv0850 was also demonstrated by Reverse transcription PCR (RT-PCR) of intergenic regions. (C) 2013 Elsevier GmbH. All rights reserved.