Abstract
Mycobacteriophage are viruses that infect mycobacteria. More than 1,400 mycobacteriophage genomes have been sequenced, coding for over one hundred thousand proteins of unknown functions. Here we investigate mycobacteriophage Giles-host protein-protein interactions (PPIs) using yeast two-hybrid screening (Y2H). A total of 25 reproducible PPIs were found for a selected set of 10 Giles proteins, including a putative virion assembly protein (gp17), the phage integrase (gp29), the endolysin (gp31), the phage repressor (gp47), and six proteins of unknown function (gp34, gp35, gp54, gp56, gp64, and gp65). We note that overexpression of the proteins is toxic to M. smegmatis, although whether this toxicity and the associated changes in cellular morphology are related to the putative interactions revealed in the Y2H screen is unclear.
Highlights
Bacteriophages are the most abundant, diverse and highly populated biological entities with an estimated 1031 phage particles in the biosphere[1]
Mycobacteriophage Giles is a temperate phage that forms stable lysogens in M. smegmatis[20]
A set of 10 Giles-encoded proteins (Table 1) were selected for screening against a M. smegmatis genomic library in a Y2H screen (Fig. 1A). These proteins represent a variety of expression and functional features, and include a putative virion assembly protein, the phage integrase, the endolysin, the phage repressor, and six proteins of unknown function
Summary
Bacteriophages are the most abundant, diverse and highly populated biological entities with an estimated 1031 phage particles in the biosphere[1]. More than 1,400 completely sequenced mycobacteriophage genomes have been described (http://phagesdb.org)[3], which have facilitated development of tools for mycobacterial genetics, and may have therapeutic potential[4] These genomes display high genetic diversity and encode an abundance of genes of unknown function[5,6]. A broad search to define Giles genes needed for lytic growth showed that more than half of the non-structural genes are dispensable for plaque formation, many show minor defects in phage production[20] These genetic and transcriptomic analysis support further analysis of mycobacteriophage Giles to elucidate protein-protein interactions[21]
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