Recognizing sweet surfaces – structure and function of a siphovirus infection device at the Gram-negative bacterial membrane

• Applicants: Professorin Dr. Stefanie Barbirz; Dr. Nina Bröker
• Subject Area: Metabolism, Biochemistry and Genetics of Microorganisms ; Structural Biology ; Virology
• Term: since 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 505655091

Bacterial viruses (bacteriophages) shuttle their genomes into bacterial cells as a key step in their infection cycle. Many bacteriophages use a tail for adsorption and DNA translocation to the host’s cytosol. Tailed phages combine core tail architectures with diversified infection parts, reflecting the various different envelope compositions encountered on their hosts. Long, non-contractile tailed siphoviruses are a predominant tailed phage type. However, the nature of the infection signal these phages receive from the bacterial envelope is not well understood and needs more investigations. Goal of the French-German project is to elucidate, with cryo-elctron microscopy (cryoEM), the first prototype structure for a glycan-specific siphovirus tail and baseplate infecting a Gram-negative host, from Salmonella model phage 9NA. This will define structural differences to phages that use protein receptors for infection. With total internal reflection fluorescence (TIRF) microscopy, and new Gram-negative model membrane set-ups, we will study siphovirus 9NA’s time-resolved genome release mechanism on a single particle level, and analyze the influence of Gram-negative membrane properties on successful infection initiation. CryoEM studies in presence of the lipopolysaccharide receptor or outer membrane vesicles will show how phage structural proteins are involved in infection initiation upon membrane contact. We will thus gain understanding of the molecular rearrangements in the siphovirus tail leading to genome release. We will moreover use these in vitro set-ups to study synergies of phage mixtures that compete for receptors on the same host and correlate this data with in vivo infection analyses on whole bacterial cells. Exploring the dynamics in bacteriophage-envelope interactions will advance our view on phage tail structure rearrangements, leading to genome release as a key step in the phage life cycle. Elucidating the tail and baseplate structure of a model system representing a widespread type of Gram-negative, glycan-specific phages will improve structure-based genome annotation and phage receptor usage prediction from sequence data. Our work will contribute substantial molecular knowledge on how phage communities behave in a given infection ecosystem, important for the development of new antibiotic treatments that employ bacteriophages.

• DFG Programme: Research Grants
• International Connection: France
• Partner Organisation: Agence Nationale de la Recherche / The French National Research Agency
• Cooperation Partners: Dr. Pascale Boulanger; Dr. Cécile Breyton