Working Paper Article Version 1 This version is not peer-reviewed

Identification of The Role of O-antigen of Nonmucoid Klebsiella pneumoniae During Bacteriophage NJS1 Infection

Version 1 : Received: 24 July 2020 / Approved: 25 July 2020 / Online: 25 July 2020 (18:55:02 CEST)

How to cite: Hao, G.; Jia, Y.; Yuan, C.; Liu, M.; Zhao, S.; Zhou, H.; Sun, S.; Wang, H. Identification of The Role of O-antigen of Nonmucoid Klebsiella pneumoniae During Bacteriophage NJS1 Infection. Preprints 2020, 2020070623 Hao, G.; Jia, Y.; Yuan, C.; Liu, M.; Zhao, S.; Zhou, H.; Sun, S.; Wang, H. Identification of The Role of O-antigen of Nonmucoid Klebsiella pneumoniae During Bacteriophage NJS1 Infection. Preprints 2020, 2020070623

Abstract

The use of bacteriophage is reemerging as a tool for combatting multi-drug resistant bacterial infections. In our previous study, we showed that colistin resistant carbapenem-resistant Klebsiella pneumoniae (ColR-CRKP) is more susceptible to killing by lytic tailed phages, including ФNJS1 specific for nonmucoid K. pneumoniae. Although we demonstrated that alteration on surface charges of ColR-CRKP promotes phage adherence and infection, the receptor for ФNJS1 was still unknown. In current study, we identified O-antigen was involved in the reversible adsorption, and outer membrane protein (OMP) FepA may be served as one of the irreversible receptors for ФNJS1. We firstly found accelerated reversible phage adsorption to ColR-CRKP cells, and that periodate treatment of bacteria inhibited the phage binding, indicating LPS may be involved in phage reversible adsorption. ФNJS1-resistant bacterial mutants screening revealed that mutants in ∆wecG(mTn5) and ∆wecA(mTn5), two genes responsible for LPS biosynthesis, affected phage adsorption capacity and phage infectivity. The loss of wzyE encoding O-antigen polymerase showed no significant difference in phage adsorption but increased phage infectivity, suggesting the long chain length of O-antigen may also be a barrier for bacteriophage infection. Among four OMP mutants including ∆fepA, ∆fhuA, ∆ompA and ∆ompC, only ∆fepA slowed phage lysis rate, suggesting FepA may be as one of irreversible receptors for ФNJS1. The results are helpful to better understand why ColR-CRKP sensitizes phage infection and to combat multi-drug resistant K. pneumoniae infections in the future.

Subject Areas

Klebsiella pneumoniae; nonmucoid; O-antigen; outer membrane protein; phage receptor

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