Involvement of the Cell Division Protein DamX in the Infection Process of Bacteriophage T4

Author:

Wenzel Sabrina1,Hess Renate1,Kiefer Dorothee1,Kuhn Andreas1ORCID

Affiliation:

1. Institute of Biology, University of Hohenheim, 190h, Garbenstr. 30, 70599 Stuttgart, Germany

Abstract

The molecular mechanism of how the infecting DNA of bacteriophage T4 passes from the capsid through the bacterial cell wall and enters the cytoplasm is essentially unknown. After adsorption, the short tail fibers of the infecting phage extend from the baseplate and trigger the contraction of the tail sheath, leading to a puncturing of the outer membrane by the tail tip needle composed of the proteins gp5.4, gp5 and gp27. To explore the events that occur in the periplasm and at the inner membrane, we constructed T4 phages that have a modified gp27 in their tail tip with a His-tag. Shortly after infection with these phages, cells were chemically cross-linked and solubilized. The cross-linked products were affinity-purified on a nickel column and the co-purified proteins were identified by mass spectrometry, and we found that predominantly the inner membrane proteins DamX, SdhA and PpiD were cross-linked. The same partner proteins were identified when purified gp27 was added to Escherichia coli spheroplasts, suggesting a direct protein–protein interaction.

Funder

Deutsche Forschungsgemeinschaft

Publisher

MDPI AG

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