Structure and Assembly of TP901-1 Virion Unveiled by Mutagenesis

Stephen R. Stockdale (1) , Barry Collins (1) , Silvia Spinelli (2) , François P. Douillard (1) , Jennifer Mahony (1) , Christian Cambillau (2) , Douwe van Sinderen (3, 1)

1 School of Microbiology
2 AFMB - Architecture et fonction des macromolécules biologiques
3 Alimentary Pharmabiot Ctr, Biosci Inst, Dept Microbiol

Bacteriophages of the Siphoviridae family represent the most abundant viral morphology in the biosphere, yet many molecular aspects of their virion structure, assembly and associated functions remain to be unveiled. In this study, we present a comprehensive mutational and molecular analysis of the temperate Lactococcus lactis-infecting phage TP901-1. Fourteen mutations located within the structural module of TP901-1 were created; twelve mutations were designed to prevent full length translation of putative proteins by nonsense mutations, while two additional mutations caused aberrant protein production. Electron microscopy and Western blot analysis of mutant virion preparations, as well as in vitro assembly of phage mutant combinations, revealed the essential nature of many of the corresponding gene products and provided information on their biological function(s). Based on the information obtained, we propose a functional and assembly model of the TP901-1 Siphoviridae virion.

Mots clés: LACTOCOCCAL BACTERIOPHAGE TP901-1; SECONDARY STRUCTURE PREDICTION; MULTIPLE SEQUENCE ALIGNMENT; PROTEIN HOMOLOGY DETECTION; TAIL-TERMINATOR PROTEIN; LACTIC-ACID BACTERIA; PHAGE-LAMBDA; CRYSTAL-STRUCTURE; BINDING PROTEIN; 3-DIMENSIONAL STRUCTURE

Domaines: Sciences du Vivant [q-bio]; Biochimie, Biologie Moléculaire

Stephen R. Stockdale, Barry Collins, Silvia Spinelli, François P. Douillard, Jennifer Mahony, et al.. Structure and Assembly of TP901-1 Virion Unveiled by Mutagenesis. PLoS ONE, 2015, pp.1-22. ⟨10.1371/journal.pone.0131676⟩. ⟨hal-01201678⟩