The Generation of an Artificial ATP Deficit Triggers Antibiotic Production in Streptomyces lividans

Abstract

In most Streptomyces species, antibiotic production is triggered in a condition of phosphate limitation, a condition that is known to be correlated with a low intracellular ATP content compared to growth in a condition of phosphate proficiency. This observation suggests that a low ATP content might be a direct trigger of antibiotic biosynthesis. In order to test this hypothesis, we introduced into the model strain Streptomyces lividans, a functional and a non-functional ATPase cloned into the replicative vector pOSV206 and expressed under the control of the strong ErmE* promoter. The functional ATPase was constituted by the α (AtpA), β (AtpB) and γ (AtpD) sub-units of the native F1 part of the ATP synthase of S. lividans that, when separated from the membrane-bound F0 part, bears an ATPase activity. The non-functional ATPase was a mutated version of the latter, bearing a 12 amino acids deletion encompassing the active site of the AtpD sub-unit. S. lividans was chosen to test our hypothesis since this strain hardly produces any antibiotics. However, it possesses the same biosynthetic pathways of various specialized metabolites as S. coelicolor, a phylogenetically closely related strain that produces these metabolites in abundance. Our results demonstrated that the over-expression of the functional ATPase, but not that of its mutated version, indeed correlated with the production of the bioactive metabolites of the CDA, RED and ACT clusters. These results confirmed the long known and mysterious link existing between a phosphate limitation leading to an ATP deficit and the triggering of antibiotic biosynthesis. Based on this work and the previous published results of our group, we propose an entirely novel conception of the nature of this link.

Metabolic adjustments in response to ATP spilling by the small DX protein in a Streptomyces strain

“…A reduced generation of ATP due to nutritional limitation in condition of high cell density or DX-mediated ATP degradation were correlated with the triggering of the production of specific antibiotics. This is consistent with previous work that demonstrated that the over-expression of an ATPase triggers antibiotic production in the weak producer Streptomyces lividans ( Seghezzi et al, 2022 ). However, even at high plating density, A37 produces higher amounts of the same specialized metabolites than A36 ( Supplementary Table S2 ).…”

Genome Analysis of a Variant of Streptomyces coelicolor M145 with High Lipid Content and Poor Ability to Synthetize Antibiotics

“…Consequently, a weaker oxidative stress could explain the lower ACT production of the TD variant compared to the original strain. Nonetheless, the TD variant still produces ACT at a low level in the condition of Pi limitation ( Figure 1 A), a condition known to promote the activation of the oxidative metabolism and thus the generation of oxidative stress [ 44 , 45 ], whereas ACT production is undetectable in the condition of Pi proficiency ( Figure 1 B). The remaining low level of ACT production in the TD variant could be due to a low level of oxidative stress due the very active fatty acids/lipids biosynthesis taking place in the TD variant.…”

Discovery and Biosynthesis of Streptolateritic Acids A–D: Acyclic Pentacarboxylic Acids from Streptomyces sp. FXJ1.172 with Promising Activity against Potato Common Scab

“…7−10 Phosphate limitation or a low intracellular ATP content causes a stress response that triggers specialized metabolite production in Streptomyces. 11,12 Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) is an oxidative phosphorylation inhibitor that inhibits the synthesis of ATP, and it has been used to induce actinorhodin production in respiratory mutants of Streptomyces coelicolor A3(2). 13 NPs assembled by type I modular polyketide synthases (PKSs) have attracted great attention due to their broad spectrum of biological activities 14 and complex biosynthetic mechanisms.…”

“…However, many of these BGCs are poorly expressed in laboratory growth conditions, making identification of the NPs they encode challenging . Activation of such “silent” BGCs enables the discovery of NPs with unprecedented structures and intriguing bioactivities. − Phosphate limitation or a low intracellular ATP content causes a stress response that triggers specialized metabolite production in Streptomyces. , Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) is an oxidative phosphorylation inhibitor that inhibits the synthesis of ATP, and it has been used to induce actinorhodin production in respiratory mutants of Streptomyces coelicolor A3(2) …”