Rufomycin Targets ClpC1 Proteolysis in Mycobacterium tuberculosis and M. abscessus.

Mary P Choules,Wei Gao,Yuehong Wang,Guido F Pauli,Jeffrey R Anderson,Chulhun L Chang,Birgit U Jaki,Seungwha Paik,Hanki Lee,Scott G Franzblau,Rui Ma,Nguyen Minh Duc,Hyun Lee,Edyta M Grzelak,Eun-Kyeong Jo,Nina M Wolf,Jinhua Cheng,James B Mcalpine,Jong Seok Lee,Joo Won Suh ,Jun Ho Choe ,Ying Jin ,Sang Hyun Cho

doi:10.1128/aac.02204-18

Abstract

ClpC1 is an emerging new target for the treatment of Mycobacterium tuberculosis infections, and several cyclic peptides (ecumicin, cyclomarin A, and lassomycin) are known to act on this target. This study identified another group of peptides, the rufomycins (RUFs), as bactericidal to M. tuberculosis through the inhibition of ClpC1 and subsequent modulation of protein degradation of intracellular proteins. Rufomycin I (RUFI) was found to be a potent and selective lead compound for both M. tuberculosis (MIC, 0.02 μM) and Mycobacterium abscessus (MIC, 0.4 μM). Spontaneously generated mutants resistant to RUFI involved seven unique single nucleotide polymorphism (SNP) mutations at three distinct codons within the N-terminal domain of clpC1 (V13, H77, and F80). RUFI also significantly decreased the proteolytic capabilities of the ClpC1/P1/P2 complex to degrade casein, while having no significant effect on the ATPase activity of ClpC1. This represents a marked difference from ecumicin, which inhibits ClpC1 proteolysis but stimulates the ATPase activity, thereby providing evidence that although these peptides share ClpC1 as a macromolecular target, their downstream effects are distinct, likely due to differences in binding.

Highlights

ClpC1 is an emerging new target for the treatment of Mycobacterium tuberculosis infections, and several cyclic peptides are known to act on this target
Strain MJM3502 was obtained by the Extract Collection of Useful Microorganisms (ECUM) at Myongji University, Republic of Korea, and was fermented in glucose-soybean starch (GSS) medium
MJM3502 was identified as a hit from the high-throughput screening (HTS) of approximately 7,000 actinomycete cultures as previously discussed [12]

Summary

Introduction

ClpC1 is an emerging new target for the treatment of Mycobacterium tuberculosis infections, and several cyclic peptides (ecumicin, cyclomarin A, and lassomycin) are known to act on this target. This study identified another group of peptides, the rufomycins (RUFs), as bactericidal to M. tuberculosis through the inhibition of ClpC1 and subsequent modulation of protein degradation of intracellular proteins. RUFI significantly decreased the proteolytic capabilities of the ClpC1/P1/P2 complex to degrade casein, while having no significant effect on the ATPase activity of ClpC1 This represents a marked difference from ecumicin, which inhibits ClpC1 proteolysis but stimulates the ATPase activity, thereby providing evidence that these peptides share ClpC1 as a macromolecular target, their downstream effects are distinct, likely due to differences in binding. ECU has been found to trigger the first proposed mechanism by uncoupling ClpC1 from ClpP activity, leading to a decrease in protein degradation

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Conclusion