Impact of Plasmid-Encoded H-NS-like Protein on blaNDM-1-Bearing IncX3 Plasmid in Escherichia coli

Abstract

Background: This study was performed to assess the role of the histone-like nucleoid-structuring (H-NS)-like protein, carried by blaNDM-1-encoding IncX3-type plasmids, in the dissemination of IncX3 plasmids.

Methods: The blaNDM-1-encoding IncX3 plasmids were analyzed using southern blot, conjugation, and competition assays. Virulence was evaluated with a Galleria mellonella infection model. An hns-knockout IncX3 plasmid was also constructed to identify the functions of plasmid-borne H-NS-like protein in Escherichia coli.

Results: The assasys detected blaNDM-1-encoding IncX3-type plasmids with similar fingerprint patterns in all New Delhi metallo-β-lactamase (NDM) 1-producing carbapenem-resistant Enterobacteriaceae. The IncX3 plasmid conferred a fitness advantage to E. coli J53 but had no effect on host virulence. Moreover, the transconjugation frequency of the hns-null IncX3 plasmid pHN330-△hns was increased by 2.5-fold compared with the wild type. This was caused by up-regulation of conjugation-related plasmid-borne genes and the partition-related gene, in the J330-pHN330-△hns strain. In addition, decreased virulence was detected with this variant.

Conclusions: Our results highlight the important role of IncX3 plasmids in the dissemination of blaNDM-1 in south China. Plasmid-encoded H-NS-like protein can inhibit plasmid conjugation, partition, and the expression of related genes, in addition to promoting virulence in the host.

Keywords: E. coli; bla NDM-1; H-NS-like protein; IncX3 plasmid; dissemination.