Viral proteins involved in the adsorption process of phage Deep-Purple infecting members of the Bacillus cereus group

Leprince, Audrey;Nuytten, Manon;Mahillon, Jacques
(2021) Bacillus ACT 2021 9th International Conference on Bacillus anthracis, cereus and thuringiensis — Location: Online (26.April.2021)

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  • Leprince, AudreyUCLouvain
    Author
  • Nuytten, ManonUCLouvain
    Author
  • Mahillon, JacquesUCLouvain
    Author
Abstract
The phage life cycle is initiated by the interaction between viral proteins and specific receptor at the bacterial surface. This adsorption step is a key determinant of the phage host spectrum and also the main target of resistance mechanisms developed by bacteria to overcome phage infection. Thus, in the light of the increase interest for phage in medical and industrial applications, it is paramount to characterize the molecular mechanisms underlying phage-host primary interactions. In this work, we were interested in identifying and characterizing the phage structural proteins involved in the adsorption of Deep-Purple, a Siphoviridae (i.e. phage possessing a long non contractile tail) infecting members of the B. cereus group [1]. Commonly, the central actors in the adsorption step are structures called Receptor Binding Proteins (RBP) which are responsible for the specific recognition of the bacterial host and subsequent triggering of genome delivery. However, recently, another type of adsorption associated protein, called evolved Distal tail (evoDit) proteins, were identified in Siphoviridae infecting Lactic Acid Bacteria (LAB). These proteins were previously thought to have only a structural role, but it appears that the presence of a carbohydrate-binding module (CBM) allows them to interact with the host surface and thus assist the RBP. Bioinformatic analysis of Deep-Purple tail protein-encoding region allowed to identify gp28 and gp29 as the putative evoDit protein and RBP, respectively. Both proteins contain a CBM, suggesting that these proteins can interact with host associated polysaccharides. When fused to a fluorescent tag, both proteins were able to decorate the bacterial surface but differences in binding spectra were observed. The RBP-gp29 only recognized strains that are sensitive to the phage whereas evoDit-gp28 bound to many strains including some that are insensitive to Deep-Purple, highlighting the fact that the evoDit is not the sole determinant of subsequent phage replication. A detailed the characterization of both RBP-gp29 and evoDit-gp28 will be presented, together with their respective involvement in Deep-Purple adsorption.
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Leprince, A., Nuytten, M., & Mahillon, J. (2021). Viral proteins involved in the adsorption process of phage Deep-Purple infecting members of the Bacillus cereus group. Bacillus ACT 2021 9th International Conference on Bacillus anthracis, cereus and thuringiensis, Online. https://hdl.handle.net/2078.5/240834