An insight into the regulation of mce4 operon of Mycobacterium tuberculosis

Nisha Rathor, Amita Chandolia, Neeraj Kumar Saini, Rajesh Sinha, Rakesh Pathak, Kushal Garima, Satendra Singh, Mandira Varma-Basil, Mridula Bose

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

The mce4 operon is reported to be involved in cholesterol utilization and intracellular survival of Mycobacterium tuberculosis (M. tuberculosis). The regulatory mechanism of this important operon was unknown so far. Here we report detection of the promoter region and regulatory factors of the mce4 operon. The in silico analyzed putative promoter region was cloned in promoter selection vector and promoter strength was measured by O-Nitrophenyl-β-d- galactopyranosidase (ONPG) assay. The transcription start site was determined by 5′ Rapid amplification of C terminal end (5′RACE). Surface stress, hypoxia and presence of cholesterol, were found to be stimulatory for mce4 operon promoter induction. Pull down assay coupled with 2D gel electrophoresis resolved many proteins; few prominent spots were processed for identification. MALDI TOF-TOF identified proteins of M. tuberculosis which supported the regulatory function of the identified promoter region and cholesterol utilization of mce4 operon. Since mce4 operon is involved in cholesterol utilization and intracellular survival of M. tuberculosis in the later phase of infection, identification of the promoter sequence as reported in the present communication may facilitate development of effective inhibitors to regulate expression of mce4 operon which may prove to be a good drug target to prevent latency in tuberculosis.

Original languageEnglish (US)
Pages (from-to)389-397
Number of pages9
JournalTuberculosis
Volume93
Issue number4
DOIs
Publication statusPublished - Jul 1 2013

    Fingerprint

Keywords

  • Mycobacterium tuberculosis mce4 Operon Regulation Stationary phase Cholesterol metabolism

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Microbiology (medical)
  • Infectious Diseases

Cite this

Rathor, N., Chandolia, A., Saini, N. K., Sinha, R., Pathak, R., Garima, K., ... Bose, M. (2013). An insight into the regulation of mce4 operon of Mycobacterium tuberculosis. Tuberculosis, 93(4), 389-397. https://doi.org/10.1016/j.tube.2013.03.007