Lactococcal Bacteriophages Require a Host Cell Wall Carbohydrate and a Plasma Membrane Protein for Adsorption and Ejection of DNA

Abstract

The mechanism of the initial steps of bacteriophage infection in Lactococcus lactis subsp. lactis C2 was investigated by using phages c2, m13, kh, 1, h, 5, and 13. All seven phages adsorbed to the same sites on the host cell wall that are composed, in part, of rhamnose. This was suggested by rhamnose inhibition of phage adsorption to cells, competition between phage c2 and the other phages for adsorption to cells, and rhamnose inhibition of lysis of phage-inoculated cultures. The adsorption to the cell wall was found to be reversible upon dilution of the cell wall-adsorbed phage. In a reaction step that apparently follows adsorption to the cell wall, all seven phages adsorbed to a host membrane protein named PIP. This was indicated by the inability of all seven phages to infect a strain selected for resistance to phage c2 and known to have a defective PIP protein. All seven phages were inactivated in vitro by membranes from wild-type cells but not by membranes from the PIP-defective, phage c2-resistant strain. The mechanism of membrane inactivation was an irreversible adsorption of the phage to PIP, as indicated by adsorption of [35S]methionine-labeled phage c2 to purified

Phages of Lactobacillus casei/paracasei: response to environmental factors and interaction with collection and commercial strains

“…The vegetative cycle of viral multiplication is initiated by phage specific adsorption to the cell wall of a susceptible bacterial host (Neve 1996). Although the adsorption process was studied for other acid lactic bacteria (Budde-Niekiel and Teuber 1987;Sijtsma et al 1988;Valyasevi et al 1990;Monteville et al 1994;Foschino et al 1995;Quiberoni and Reinheimer 1998;Binetti et al 2002;Lu et al 2003;Quiberoni et al 2004) the information available on Lact. casei/paracasei is still scarce since it was only reported for system /PL-1/Lact.…”

Loss of function mutations inepaRconfer resistance to phage NPV1 infection inEnterococcus faecalisOG1RF

“…Phage receptors have been extensively studied in certain phage families, including the T series phages, Mu, and λ for Gram-negative bacteria (8-11). Some phage receptors have been characterized in Gram-positive bacteria, including receptors for ϕSPP-1 of Bacillus subtilis (12) and the phage c2 group of Lactococcus lactis (13, 14). YueB, the ϕSPP-1 receptor, and PIP, the phage c2 receptor, are orthologs and are required for irreversible phage adsorption (12).…”

Temperate phages TP901-1 and ϕLC3, belonging to the P335 species, apparently use different pathways for DNA injection inLactococcus lactissubsp.cremoris3107

“…In a number of lactococcal phage/host systems, phage adsorption has been shown to involve attachment of the phage to a carbohydrate moiety in the cell wall, such as galactose and/or rhamnose and to a lesser extent glucose (Keogh & Pettingill, 1983;Valyasevi et al, 1990Valyasevi et al, , 1995Monteville et al, 1994). For phage c2, the primary adsorption step was found to be reversible, whereas the following binding to a receptor in the cell membrane was found to be irreversible (Valyasevi et al, 1991;Monteville et al, 1994).…”

“…In a number of lactococcal phage/host systems, phage adsorption has been shown to involve attachment of the phage to a carbohydrate moiety in the cell wall, such as galactose and/or rhamnose and to a lesser extent glucose (Keogh & Pettingill, 1983;Valyasevi et al, 1990Valyasevi et al, , 1995Monteville et al, 1994). For phage c2, the primary adsorption step was found to be reversible, whereas the following binding to a receptor in the cell membrane was found to be irreversible (Valyasevi et al, 1991;Monteville et al, 1994). Mutation analysis in IL1403 indicated that a polysaccharide may be involved in the adsorption of phage bIL170 and j635, both belonging to the 936 phage species (Dupont et al, 2004a) while the receptor binding protein was identified to be ORF20 and ORF18 in phage bIL170 and sk1, respectively (Dupont et al, 2004b).…”