A dimeric holin/antiholin complex controls lysis by phage T4

verfasst von

Jan Michel Frederik Schwarzkopf, Denise Mehner-Breitfeld, Thomas Brüser

Abstract

Lytic phages control the timepoint of host cell lysis by timing the holin-mediated release of cell wall-degrading endolysins. In phage T4, the antiholin RI inhibits the holin T, thereby preventing the early release of the T4 endolysin and lysis. The antiholin achieves lysis inhibition (LIN) in response to phage superinfections, thereby increasing the chance for lysis in an environment with a lower phage concentration. The holin T consists of a small N-terminal cytoplasmic domain, a transmembrane helix, and a periplasmic C-terminal domain. The antiholin is targeted to the periplasm by a cleavable signal peptide. Recently, the periplasmic soluble domains of the holin and the antiholin were found to form T2/RI2 tetramers in crystals. To investigate the functional relevance of this complex, we reconstituted LIN in a phage-free system, using only RI, T, and endolysin, and combined targeted mutagenesis with functional analyses. Inactivation of the RI signal peptide cleavage site did not abolish LIN, indicating that RI can function in a membrane-bound state, which argued against the tetramer. This led to analyses showing that only one of the two T/RI interfaces in the tetramer is physiologically relevant, which is also the only interaction site predicted by AlphaFold2. Some holin mutations at this interaction site prevented lysis, suggesting that the RI interaction likely acts by blocking the holin oligomerization required for hole formation. We conclude that LIN is mediated by a dimeric T/RI complex that, unlike the tetramer, can be easily formed when both partners are membrane-anchored.

Organisationseinheit(en)

Institut für Mikrobiologie

Typ

Artikel

Journal

Frontiers in Microbiology

Band

15

ISSN

1664-302X

Publikationsdatum

05.09.2024

Publikationsstatus

Veröffentlicht

Peer-reviewed

Ja

ASJC Scopus Sachgebiete

Allgemeine Biochemie, Genetik und Molekularbiologie, Mikrobiologie (medizinisch), Mikrobiologie

Fachgebiet (basierend auf ÖFOS 2012)

Molekularbiologie, Mikrobiologie, Strukturbiologie

Elektronische Version(en)

https://doi.org/10.3389/fmicb.2024.1419106 (Zugang: Offen)