Targeting PknL as a novel therapeutic strategy against Mycobacterium tuberculosis

Abstract

Abstract: INTRODUCTION: The bacterium “Mycobacterium tuberculosis (MTB)” causes tuberculosis, which is currently regarded as the most common cause of bacterial infection-related death globally. In addition to eukaryotes, MTB also encodes 11 “serine/threonine protein kinases (STPKs),” including PknA, B, C, D, E, F, G, H, J, K, and L that control cell division, cell development, metabolism, and dormancy. MATERIALS AND METHODS: The antisense and sense-orientated PknL have been cloned utilizing the shuttle mycobacterial vector-pAZI9018b, which is IPTG inducible. Utilizing the appropriate primers, the PknL (Rv2176) full-length gene from MTB has been amplified. The product which was amplified was then inoculated inside the vector at the “5’BamH1 and 3’Nde1” regions to produce PknL in the sense orientation (PknL-S) resembling PknL-S, and likewise, amplified substance was introduced into the vector at the 3’BamHI and 5’NdeI sites for producing PknL in the antisense orientation (PknL-AS). RESULTS: Regardless of the absence or presence of an inducer, PknL-AS had lower transcription levels than Wild type and PknL-S, but PknL-AS revealed a significant decrease in PknL expression during the end of 2nd week with 1 mM of IPTG and a further decrease on day 10 with 10 mM IPTG. In the presence of 10 mM IPTG, PknL expression has been close to, 1.8 log in Wild type, 1.4 log in PknL-AS, and 2.4 log in PknL-S. CONCLUSION: It has been clear that PknL may read environmental cues and respond appropriately, assisting bacteria to adapt to stressful situations in the host by lowering bacterial development, leading to a dormant state, and allowing bacteria to stay inside the host.