TY - JOUR

T1 - Induction of proteome changes involved in the cloning of mcr-1 and mcr-2 genes in Escherichia coli DH5-α strain to evaluate colistin resistance

AU - Feizi, Hadi

AU - Alizadeh, Maryam

AU - Azimi, Hadi

AU - Khodadadi, Ehsaneh

AU - Kamounah, Fadhil S.

AU - Ganbarov, Khudaverdi

AU - Ghotaslou, Reza

AU - Rezaee, Mohammad Ahangarzadeh

AU - Kafil, Hossein Samadi

N1 - Funding Information: Funding: This study was supported by the Drug Applied Research Centre , Tabriz University of Medical Sciences , with grant number 64650 . All steps were done according to the Helsinki Declaration. Publisher Copyright: © 2023 The Author(s)

PY - 2024

Y1 - 2024

N2 - Objectives: Plasmid genes, termed mobile colistin resistance-1 (mcr-1) and mobile colistin resistance-2 (mcr-2), are associated with resistance to colistin in Escherichia coli (E. coli). These mcr genes result in a range of protein modifications contributing to colistin resistance. This study aims to discern the proteomic characteristics of E. coli–carrying mcr-1 and mcr-2 genes. Furthermore, it evaluates the expression levels of various proteins under different conditions (with and without colistin). Methods: Plasmid extraction was performed using an alkaline lysis–based plasmid extraction kit, whereas polymerase chain reaction was used to detect the presence of mcr-1 and mcr-2 plasmids. The E. coli DH5α strain served as the competent cell for accepting and transforming mcr-1 and mcr-2 plasmids. We assessed proteomic alterations in the E. coli DH5α strain both with and without colistin in the growth medium. Proteomic data were analysed using mass spectrometry. Results: The findings revealed significant protein changes in the E. coli DH5α strain following cloning of mcr-1 and mcr-2 plasmids. Of the 20 proteins in the DH5α strain, expression in 8 was suppressed following transformation. In the presence of colistin in the culture medium, 39 new proteins were expressed following transformation with mcr-1 and mcr-2 plasmids. The proteins with altered expression play various roles. Conclusion: The results of this study highlight numerous protein alterations in E. coli resulting from mcr-1 and mcr-2–mediated resistance to colistin. This understanding can shed light on the resistance mechanism. Additionally, the proteomic variations observed in the presence and absence of colistin might indicate potential adverse effects of indiscriminate antibiotic exposure on treatment efficacy and heightened pathogenicity of microorganisms.

AB - Objectives: Plasmid genes, termed mobile colistin resistance-1 (mcr-1) and mobile colistin resistance-2 (mcr-2), are associated with resistance to colistin in Escherichia coli (E. coli). These mcr genes result in a range of protein modifications contributing to colistin resistance. This study aims to discern the proteomic characteristics of E. coli–carrying mcr-1 and mcr-2 genes. Furthermore, it evaluates the expression levels of various proteins under different conditions (with and without colistin). Methods: Plasmid extraction was performed using an alkaline lysis–based plasmid extraction kit, whereas polymerase chain reaction was used to detect the presence of mcr-1 and mcr-2 plasmids. The E. coli DH5α strain served as the competent cell for accepting and transforming mcr-1 and mcr-2 plasmids. We assessed proteomic alterations in the E. coli DH5α strain both with and without colistin in the growth medium. Proteomic data were analysed using mass spectrometry. Results: The findings revealed significant protein changes in the E. coli DH5α strain following cloning of mcr-1 and mcr-2 plasmids. Of the 20 proteins in the DH5α strain, expression in 8 was suppressed following transformation. In the presence of colistin in the culture medium, 39 new proteins were expressed following transformation with mcr-1 and mcr-2 plasmids. The proteins with altered expression play various roles. Conclusion: The results of this study highlight numerous protein alterations in E. coli resulting from mcr-1 and mcr-2–mediated resistance to colistin. This understanding can shed light on the resistance mechanism. Additionally, the proteomic variations observed in the presence and absence of colistin might indicate potential adverse effects of indiscriminate antibiotic exposure on treatment efficacy and heightened pathogenicity of microorganisms.

KW - AMR

KW - Colistin

KW - E. coli

KW - MALDI-TOF MS

KW - Mcr-1

KW - Mcr-2

U2 - 10.1016/j.jgar.2023.12.018

DO - 10.1016/j.jgar.2023.12.018

M3 - Journal article

C2 - 38154746

AN - SCOPUS:85182645572

VL - 36

SP - 151

EP - 159

JO - Journal of Global Antimicrobial Resistance

JF - Journal of Global Antimicrobial Resistance

SN - 2213-7165

ER -