A Halocin Promotes DNA Uptake in Haloferax mediterranei.

Shaoxing Chen,Jingwen Liu,Siqi Sun,Hua Xiang,Gregory A Korfanty

doi:10.3389/fmicb.2019.01960

Abstract

Halocins are antimicrobial peptides or proteins that are produced by halophilic archaea. Although their function in inhibiting the growth of closely related haloarchaeal strains is well known, other physiological functions of halocins have also been proposed in recent years. To unveil the possible function and mechanism of halocins in DNA uptake, the halocin H4 producing strain Haloferax mediterranei DF50-ΔEPS (incapable of EPS production) was used in this study. We found that deletion of the halH4 resulted in the strain DF50-ΔEPSΔhalH4 which exhibited loss of natural DNA uptake ability. Moreover, supernatants of the halocin producing strain were capable of inducing the ability to uptake DNA. Obviously, halocin is likely responsible for inducing DNA uptake. Cell surface ultrastructures of these strains are varied from strains DF50-ΔEPS to DF50-ΔEPSΔhalH4. The cell surface of strain DF50-ΔEPS is rough due to numerous pinholes, while that of the strain DF50-ΔEPSΔhalH4 is smooth without visible pinholes. The morphology of the halH4 complemented strain, DF50-ΔEPSΔhalH4::H4, shows an intermediate phenotype between strains DF50-ΔEPS and DF50-ΔEPSΔhalH4. We speculate that halocin H4 may accelerate DNA uptake by perforating the cell surface ultrastructure. The halocin H4 may represent a novel inducer or activator of DNA uptake in Hfx. mediterranei.

Highlights

Archaeocins are antimicrobial peptides or proteins produced by archaea, currently including the halocins produced by halophilic archaea as well as sulfolobicins by Sulfolobus spp. (O’Connor and Shand, 2002)
The halH4 gene was deleted from strain DF50- EPS resulting in strain DF50- EPS halH4, which was verified by PCR amplification (Supplementary Figure S1)
Considering the overlap of the reported natural competence formation period, cells of Hfx. mediterranei at late exponential phase or at early stationary phase have been used for performing the simulation of DNA uptake in haloarchaea

Summary

Introduction

Archaeocins are antimicrobial peptides or proteins produced by archaea, currently including the halocins produced by halophilic archaea as well as sulfolobicins by Sulfolobus spp. (O’Connor and Shand, 2002). 11 halocins have been reported, i.e., HalHA1, HalHA3, HalA4, HalH1, HalH4, HalH6/H7, HalR1, HalSech7A, HalSH10, HalS8, and HalC8 (O’Connor and Shand, 2002; Li et al, 2003; Pasicet al., 2008; Karthikeyan et al, 2013; Besse et al, 2015; Kumar et al, 2016; Kumar and Tiwari, 2017; Mazguene et al, 2017). Among these halocins, the genes encoding H6/H7, H4, S8, and C8 have been described and analyzed in depth as well (Meseguer et al, 1995; Cheung et al, 1997; Sun et al, 2005). It would be interesting to investigate other possible functions of halocins in the environment, e.g., their involvement in genetic exchange or natural transformation

Methods

Results

Conclusion