A synthetic diterpene analogue inhibits mycobacterial persistence and biofilm formation by targeting (p)ppGpp synthetases.
Alexander G Tkachenko, Natalya M Kashevarova, Roman Yu Sidorov, Larisa Yu Nesterova, Anna V Akhova, Ivan V Tsyganov, Vladimir Yu Vaganov, Sergei A Shipilovskikh, Aleksandr E Rubtsov, Andrei V Malkov
Author Information
Alexander G Tkachenko: Laboratory of Microbial Adaptation, Institute of Ecology and Genetics of Microorganisms, Perm Federal Research Center, Russian Academy of Sciences, Ural Branch, Goleva 13, Perm, 614081, Perm Krai, Russia; Perm State University, Bukireva 15, Perm, 614990, Perm Krai, Russia. Electronic address: agtkachenko@iegm.ru.
Natalya M Kashevarova: Laboratory of Microbial Adaptation, Institute of Ecology and Genetics of Microorganisms, Perm Federal Research Center, Russian Academy of Sciences, Ural Branch, Goleva 13, Perm, 614081, Perm Krai, Russia.
Roman Yu Sidorov: Laboratory of Microbial Adaptation, Institute of Ecology and Genetics of Microorganisms, Perm Federal Research Center, Russian Academy of Sciences, Ural Branch, Goleva 13, Perm, 614081, Perm Krai, Russia; Perm State University, Bukireva 15, Perm, 614990, Perm Krai, Russia.
Larisa Yu Nesterova: Laboratory of Microbial Adaptation, Institute of Ecology and Genetics of Microorganisms, Perm Federal Research Center, Russian Academy of Sciences, Ural Branch, Goleva 13, Perm, 614081, Perm Krai, Russia; Perm State University, Bukireva 15, Perm, 614990, Perm Krai, Russia.
Anna V Akhova: Laboratory of Microbial Adaptation, Institute of Ecology and Genetics of Microorganisms, Perm Federal Research Center, Russian Academy of Sciences, Ural Branch, Goleva 13, Perm, 614081, Perm Krai, Russia; Perm State University, Bukireva 15, Perm, 614990, Perm Krai, Russia.
Ivan V Tsyganov: Laboratory of Microbial Adaptation, Institute of Ecology and Genetics of Microorganisms, Perm Federal Research Center, Russian Academy of Sciences, Ural Branch, Goleva 13, Perm, 614081, Perm Krai, Russia; Perm State University, Bukireva 15, Perm, 614990, Perm Krai, Russia.
Vladimir Yu Vaganov: Perm State University, Bukireva 15, Perm, 614990, Perm Krai, Russia.
Sergei A Shipilovskikh: Perm State University, Bukireva 15, Perm, 614990, Perm Krai, Russia.
Aleksandr E Rubtsov: Perm State University, Bukireva 15, Perm, 614990, Perm Krai, Russia. Electronic address: rubtsov@psu.ru.
Andrei V Malkov: Department of Chemistry, Loughborough University Address: University Road, Loughborough, Leicestershire LE11 3TU, UK. Electronic address: a.malkov@lboro.ac.uk.
Bacterial persistence coupled with biofilm formation is directly associated with failure of antibiotic treatment of tuberculosis. We have now identified 4-(4,7-DiMethyl-1,2,3,4-tetrahydroNaphthalene-1-yl)Pentanoic acid (DMNP), a synthetic diterpene analogue, as a lead compound that was capable of suppressing persistence and eradicating biofilms in Mycobacterium smegmatis. By using two reciprocal experimental approaches - Δrel and ΔrelZ gene knockout mutations versus rel and relZ overexpression technique - we showed that both Rel and RelZ (p)ppGpp synthetases are plausible candidates for serving as targets for DMNP. In vitro, DMNP inhibited (p)ppGpp-synthesizing activity of purified Rel in a concentration-dependent manner. These findings, supplemented by molecular docking simulation, suggest that DMNP targets the structural sites shared by Rel, RelZ, and presumably by a few others as yet unidentified (p)ppGpp producers, thereby inhibiting persister cell formation and eradicating biofilms. Therefore, DMNP may serve as a promising lead for development of antimycobacterial drugs.
