Koběrská Markéta, Veselá Ludmila, Novotná Michaela, Mahor Durga, Mazumdar Aninda, Pinďáková Nikola, Omena Petravicius Pamela, Pokorná Julie, Kameník Zdeněk, Balíková Novotná Gabriela
Institute of Microbiology of the CAS; BIOCEV, Vestec, Czech Republic.
Department of Genetics and Microbiology, Faculty of Science, Charles University, Prague, Czech Republic.
mBio. 2025 Sep 10;16(9):e0156825. doi: 10.1128/mbio.01568-25. Epub 2025 Aug 12.
ABCF-ATPases are increasingly recognized as translation factors that rescue stalled ribosomes when they encounter difficult mRNA templates or are stalled by antibiotics. The latter defines antibiotic resistance ABCF (ARE ABCF) proteins, known for their role in antibiotic resistance. However, in this study, we reveal a broader role of ARE ABCFs in antibiotic-responsive regulation. Using genetic, OMICs, and biochemical approaches, we showed that ARE ABCF proteins TiaA and Are5sc in use their resistance functions to modulate specialized metabolism and proteosynthesis in response to lincosamide, streptogramin A, and pleuromutilin (LSP) antibiotics. Although under LSP exposure, either Are5sc or TiaA is essential for activating the biosynthesis of the redox-active antimicrobial actinorhodin, these proteins exhibit distinct functions at the proteome level, defined by their resistance profiles and temporally regulated expression. Are5sc facilitates early adaptive responses by modulating the WblC regulon across a broad range of LSP concentrations, while TiaA is induced later, specifically at higher concentrations, where it suppresses antibiotic stress responses, particularly against pleuromutilins. TiaA function thus reflects the ecological context of LSP antibiotics as pleuromutilins are produced by fungi, whereas lincosamides/streptogramins originate from actinomycetes. Our findings demonstrate that ARE ABCF proteins, through their resistance function, act as global regulators of translation, mirroring the roles of non-ARE ABCF proteins like EttA. This highlights their broader ecological and physiological significance, extending beyond their established role in antibiotic resistance.
Bacteria adapt to diverse stimuli mainly through transcriptional changes that regulate adaptive protein factors. Here, we show that responses to protein synthesis-inhibiting antibiotics are fine-tuned by antibiotic resistance ABCF proteins at the translational level, enabling bacteria to differentiate between antibiotic classes and concentrations for a tailored response. Additionally, we have demonstrated that these proteins can specialize in conferring high-level resistance to specific antibiotics. Given their prevalence in pathogenic bacteria, antibiotic resistance ABCF (ARE ABCF) proteins may play a crucial role in resistance development, particularly against new antibiotics targeting the ribosomal catalytic center, presenting a significant challenge for antimicrobial therapy.
ABCF - ATP酶越来越被认为是一种翻译因子，当核糖体遇到困难的mRNA模板或被抗生素阻滞时，它能拯救停滞的核糖体。后者定义了抗生素抗性ABCF（ARE ABCF）蛋白，其因在抗生素抗性中的作用而闻名。然而，在本研究中，我们揭示了ARE ABCF在抗生素反应调节中更广泛的作用。通过遗传学、组学和生化方法，我们表明ARE ABCF蛋白TiaA和Are5sc利用它们的抗性功能来调节特殊代谢和蛋白质合成，以响应林可酰胺、链阳菌素A和截短侧耳素（LSP）类抗生素。尽管在LSP暴露下，Are5sc或TiaA对于激活具有氧化还原活性的抗菌放线紫红素的生物合成至关重要，但这些蛋白在蛋白质组水平上表现出不同的功能，这由它们的抗性谱和时间调控表达所定义。Are5sc通过在广泛的LSP浓度范围内调节WblC调节子来促进早期适应性反应，而TiaA在后期被诱导，特别是在较高浓度下，它抑制抗生素应激反应，尤其是针对截短侧耳素的反应。因此，TiaA的功能反映了LSP抗生素的生态背景，因为截短侧耳素由真菌产生，而林可酰胺/链阳菌素源自放线菌。我们的研究结果表明，ARE ABCF蛋白通过其抗性功能，作为翻译的全局调节因子，类似于非ARE ABCF蛋白如EttA的作用。这突出了它们更广泛的生态和生理意义，超出了它们在抗生素抗性中已确立的作用。
细菌主要通过调节适应性蛋白因子的转录变化来适应各种刺激。在这里，我们表明对抗生素抑制蛋白质合成的反应在翻译水平上由抗生素抗性ABCF蛋白进行微调，使细菌能够区分抗生素类别和浓度以进行定制反应。此外，我们已经证明这些蛋白可以专门赋予对特定抗生素的高水平抗性。鉴于它们在病原菌中普遍存在，抗生素抗性ABCF（ARE ABCF）蛋白可能在抗性发展中起关键作用，特别是针对靶向核糖体催化中心的新型抗生素，这对抗菌治疗提出了重大挑战。
