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Upregulation of the clpB gene in response to heat shock and beta-lactam antibiotics in Acinetobacter baumannii

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Abstract

The role of the clpB gene encoding HSP/chaperone ClpB was evaluated in the multiresistant antibiotic cells of Acinetobacter baumannii (RS4 strain) under stress-induced heat shock and different beta-lactams. The expression of the clpB gene was assessed by qPCR during heat shock at 45 °C and subinhibitory concentrations of ampicillin (30 μg mL−1), amoxicillin + sulbactam (8/12 μg mL−1), cefepime (30 μg mL−1), sulfamethoxazole + trimethoprim (120/8 μg mL−1) and meropenem (18 μg mL−1). The results indicated a transient increase in clpB transcription in all treatments except cefepime. Both in the presence of ampicillin and amoxicillin/sulbactam for 20 min, the mRNA-clpB synthesis was 1.4 times higher than that of the control at time zero. Surprisingly, the mRNA-clpB levels were more than 30-fold higher after 10 min of incubation with meropenem and more than eightfold higher in the presence of trimethoprim/sulfamethoxazole. In addition, western blot assays showed that the RS4 strain treated with meropenem showed a marked increase in ClpB protein expression. Our data indicate that during exposure to beta-lactams, A. baumannii adjusts the transcription levels of the clpB mRNA and protein to respond to stress, suggesting that the chaperone may act as a key cellular component in the presence of antibiotics in this bacterium.

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Funding

W. Y. Lazaretti is a professional funded by the Brazilian National Health Surveillance Agency (ANVISA). E. L. Santos was a fellow of the Brazilian National Council for Scientific and Technological Development (CNPq).

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Authors and Affiliations

  1. Laboratório de Bioquímica Molecular, Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, UNIOESTE, Rua Universitária, 2069, Cascavel, PR, 85814-110, Brazil

    Waleska Yana Lazaretti, Elaine Luzia dos Santos, José Luis da-Conceição Silva, Marina Kimiko Kadowaki, Alexandre Maller & Rita de Cássia Garcia Simão

  2. Laboratório de Micologia Clínica, Hospital Universitário do Oeste do Paraná, Universidade Estadual do Oeste do Paraná, UNIOESTE, Cascavel, PR, Brazil

    Rinaldo Ferreira Gandra

Authors
  1. Waleska Yana Lazaretti
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  2. Elaine Luzia dos Santos
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  3. José Luis da-Conceição Silva
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  4. Marina Kimiko Kadowaki
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  5. Rinaldo Ferreira Gandra
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  7. Rita de Cássia Garcia Simão
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Correspondence to Rita de Cássia Garcia Simão.

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11033_2019_5209_MOESM1_ESM.pptx

Supplementary material 1 (PPTX 3013 kb). Fig. S1. Agarose gel electrophoresis showing the products of RNA extraction obtained from the heat shock treatments and exposure to antibiotics of the multidrug resistant A. baumannii RS4 strain. (A) Heat shock at 45 °C, (B) meropenem, (C) cefepime, (D) sulfamethoxazole + trimethoprim, (E) ampicillin, (F) amoxicillin + sulbactam. (T0) time zero, (T10) 10 min, (T20) 20 min and (T30) 30 min

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Lazaretti, W.Y., dos Santos, E.L., da-Conceição Silva, J.L. et al. Upregulation of the clpB gene in response to heat shock and beta-lactam antibiotics in Acinetobacter baumannii. Mol Biol Rep 47, 1499–1505 (2020). https://doi.org/10.1007/s11033-019-05209-4

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