Setting Up a Better Infection: Overexpression of the Early Bacteriophage T4 Gene motB During Infection Results in a More Favorable tRNA Pool for the Phage.

Abstract

Background: : Although many bacteriophage T4 early genes are nonessential with unknown functions, they are believed to aid in the takeover of the Escherichia coli host. Understanding the functions of these genes could be helpful to develop novel antibacterial strategies. MotB, encoded by a previously uncharacterized T4 early gene, is a DNA-binding protein that compacts the host nucleoid and alters host gene expression.

Methods: : MotB structure was predicted by AlphaFold 2. RNA-seq and mass spectrometry (MS) analyses were performed to determine RNA and protein changes when motB was overexpressed in E. coli BL21(DE3) ±5 min T4 infection.

Results: : MotB structure is predicted to be a two-domain protein with N-terminal Kyprides-Onzonis-Woese and C-terminal oligonucleotide/oligosaccharide-fold domains. In E. coli B, motB overexpression during infection does not affect T4 RNAs, but affects the expression of host genes, including the downregulation of 21 of the 84 chargeable host tRNAs. Many of these tRNAs are used less frequently by T4 or have a counterpart encoded within the T4 genome. The MS analyses indicate that the levels of multiple T4 proteins are changed by motB overexpression.

Conclusion: : Our results suggest that in this E. coli B host, motB is involved in establishing a more favorable tRNA pool for the phage during infection.