Overexpression of the Bacteriophage T4 motB Gene Alters H-NS Dependent Repression of Specific Host DNA

• Published in: Viruses Vol. 13; no. 1; p. 84
• Main Authors: Patterson-West, Jennifer, Tai, Chin-Hsien, Son, Bokyung, Hsieh, Meng-Lun, Iben, James R, Hinton, Deborah M
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 09.01.2021; MDPI
• Subjects: Bacteria - metabolism; Bacteria - virology; Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; bacteriophage T4; Bacteriophage T4 - genetics; Base Sequence; Conserved sequence; Deoxyribonuclease; Deoxyribonucleic acid; DNA; DNA-binding protein; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - metabolism; E coli; Footprinting; Gene Expression; Genes, Viral; Glycerol; H-NS; Homology; host takeover; Host-Pathogen Interactions; Kinases; MotB; nucleoid; Oligonucleotides; Oligosaccharides; Phages; Phylogeny; Potassium; Proteins; Ribonucleic acid; RNA; Structure-function relationships; T-Phages - genetics; Taxonomy; Transcriptomes; United States > US; host takeover; DNA-binding protein; MotB; RNA-seq; H-NS; transcriptome analysis; nucleoid; bacteriophage T4
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v13010084

The bacteriophage T4 early gene product MotB binds tightly but nonspecifically to DNA, copurifies with the host Nucleoid Associated Protein (NAP) H-NS in the presence of DNA and improves T4 fitness. However, the T4 transcriptome is not significantly affected by a knockdown. Here we have investigated the phylogeny of MotB and its predicted domains, how MotB and H-NS together interact with DNA, and how heterologous overexpression of impacts host gene expression. We find that is highly conserved among . Although the MotB sequence has no homology to proteins of known function, predicted structure homology searches suggest that MotB is composed of an N-terminal Kyprides-Onzonis-Woese (KOW) motif and a C-terminal DNA-binding domain of oligonucleotide/oligosaccharide (OB)-fold; either of which could provide MotB's ability to bind DNA. DNase I footprinting demonstrates that MotB dramatically alters the interaction of H-NS with DNA in vitro. RNA-seq analyses indicate that expression of plasmid-borne up-regulates 75 host genes; no host genes are down-regulated. Approximately 1/3 of the up-regulated genes have previously been shown to be part of the H-NS regulon. Our results indicate that MotB provides a conserved function for and suggest a model in which MotB functions to alter the host transcriptome, possibly by changing the association of H-NS with the host DNA, which then leads to conditions that are more favorable for infection.