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. 2022 Dec 9:15:7227-7234.
doi: 10.2147/IDR.S390765. eCollection 2022.

IS 26-Mediated Formation of a Hybrid Plasmid Carrying mcr-1.1

Renjie Wu #  1   2   3 , Luchao Lv #  1   2   3 , Chengzhen Wang  1   2   3 , Guolong Gao  1   2   3 , Kaiyang Yu  1   2   3 , Zhongpeng Cai  1   2   3 , Yiyun Liu  1   2   3 , Jun Yang  1   2   3 , Jian-Hua Liu  1   2   3   4
Affiliations

IS 26-Mediated Formation of a Hybrid Plasmid Carrying mcr-1.1

Renjie Wu et al. Infect Drug Resist. .

Abstract

Purpose: The objective of this study was to elucidate the characteristics and mechanism of formation of the fusion plasmid pHNSHP24 carrying mcr-1.1.

Materials and methods: mcr-1.1-bearing Escherichia coli SHP24 and the corresponding transconjugant were subjected to whole-genome sequencing (WGS) combining the Illumina and MinION platforms to obtain the complete sequences of the fusion plasmid and its original plasmids.

Results: Complete sequence analysis and S1 nuclease-pulsed field gel electrophoresis (S1-PFGE) results indicated that E. coli SHP24 carried four plasmids: mcr-1.1-harboring phage-like plasmid pHNSHP24-3, F53:A-:B- plasmid pHNSHP24-4, pHNSHP24-1, and pHNSHP24-2. However, the plasmid pHNSHP24 carrying mcr-1.1 presents in the transconjugant differed from the four plasmids in the donor strain SHP24. Further analysis showed that pHNSHP24 may be the fusion product of pHNSHP24-3 and pHNSHP24-4 and is formed through a replicative transposition mechanism mediated by IS26 in E. coli SHP24.

Conclusion: This study is the first to report the fusion of an mcr-1.1-harboring phage-like pO111 plasmid and an F53:A-:B- plasmid mediated by IS26. Our findings revealed the role of phage-like and fusion plasmids in the dissemination of mcr-1.1.

Keywords: IS26; fusion plasmid; mcr-1.1; pO111 plasmid.

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Conflict of interest statement

The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
Complete sequence comparison of pO111-IncFII plasmid pHNSHP24 with phage-like plasmid, IncFII plasmid, Escherichia phage P1 (AF234172), and Escherichia phage P7 (AF503408). pMCR-1-P3 (KX880944), pO111_2 (AP010962), pMCR_SCKP-LL83 (MF510496), pPC6-mcr1 (CP080254), and pD72C (MK419152) are phage-like plasmids. pCAU16175_3 (CP047381) and pCP8-3-IncFII (CP053737) are IncFII plasmids. Functions encoded by different genes are represented in different colors, as shown in the square.
Figure 2
Figure 2
The proposed mechanism of the plasmid fusion. (A) Complete sequence comparison of fusion plasmid pHNSHP24 with daughter plasmids pHNSHP24-3 and pHNSHP24-4. Functions encoded by different genes are represented in different colors, as exhibited in the square. (B) A proposed model for the IS26-mediated formation of the fusion plasmid. Green lines and annulus represent donor plasmid IncF53:A-:B-. Blue lines and annulus represent target plasmid pO111. Red arrows represent the resistance genes. Yellow arrows represent the mobile element. Pink arrows represent the 8-bp target site (ATCGCTTA). Dark grey arrows labeled numbers 1 and 2 represent recombinase family gene (named refP) and autotransporter outer membrane gene (named aomP), respectively. Light grey arrows labeled numbers 3 and 4 represent phage-associated genes. Blue arrows represent the two pairs of hybrid primers spanning the regions of the fusion plasmid. The IS26 in plasmid pHNSHP24-4 (F53:A-:B-) attached the TSD in plasmid pHNSHP24-3 (pO1111), causing the formation of a Shapiro intermediate. DNA replication at the intermediate branch generated a cointegrate containing a duplication of IS26 and 8-bp TSD.
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