Overexpression of the Bacteriophage T4 <i>motB</i> Gene Alters H-NS Dependent Repression of Specific Host DNA
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 <i>motB</i> knockdown. H...
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| Format: | Article |
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MDPI AG
2021-01-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/13/1/84 |
| _version_ | 1797413860549853184 |
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| author | Jennifer Patterson-West Chin-Hsien Tai Bokyung Son Meng-Lun Hsieh James R. Iben Deborah M. Hinton |
| author_facet | Jennifer Patterson-West Chin-Hsien Tai Bokyung Son Meng-Lun Hsieh James R. Iben Deborah M. Hinton |
| author_sort | Jennifer Patterson-West |
| collection | DOAJ |
| description | 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 <i>motB</i> 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 <i>motB</i> impacts host gene expression. We find that <i>motB</i> is highly conserved among <i>Tevenvirinae</i>. 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 <i>motB</i> 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 <i>Tevenvirinae</i> 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. |
| first_indexed | 2024-03-09T05:24:08Z |
| format | Article |
| id | doaj.art-3f689506da654c73b58c9479459c2afc |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T05:24:08Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-3f689506da654c73b58c9479459c2afc2023-12-03T12:37:52ZengMDPI AGViruses1999-49152021-01-011318410.3390/v13010084Overexpression of the Bacteriophage T4 <i>motB</i> Gene Alters H-NS Dependent Repression of Specific Host DNAJennifer Patterson-West0Chin-Hsien Tai1Bokyung Son2Meng-Lun Hsieh3James R. Iben4Deborah M. Hinton5Gene Expression and Regulation Section, Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USACenter for Cancer Research, Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USAGene Expression and Regulation Section, Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USAGene Expression and Regulation Section, Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USAMolecular Genomics Core, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USAGene Expression and Regulation Section, Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USAThe 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 <i>motB</i> 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 <i>motB</i> impacts host gene expression. We find that <i>motB</i> is highly conserved among <i>Tevenvirinae</i>. 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 <i>motB</i> 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 <i>Tevenvirinae</i> 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.https://www.mdpi.com/1999-4915/13/1/84bacteriophage T4MotBH-NSnucleoidhost takeoverDNA-binding protein |
| spellingShingle | Jennifer Patterson-West Chin-Hsien Tai Bokyung Son Meng-Lun Hsieh James R. Iben Deborah M. Hinton Overexpression of the Bacteriophage T4 <i>motB</i> Gene Alters H-NS Dependent Repression of Specific Host DNA Viruses bacteriophage T4 MotB H-NS nucleoid host takeover DNA-binding protein |
| title | Overexpression of the Bacteriophage T4 <i>motB</i> Gene Alters H-NS Dependent Repression of Specific Host DNA |
| title_full | Overexpression of the Bacteriophage T4 <i>motB</i> Gene Alters H-NS Dependent Repression of Specific Host DNA |
| title_fullStr | Overexpression of the Bacteriophage T4 <i>motB</i> Gene Alters H-NS Dependent Repression of Specific Host DNA |
| title_full_unstemmed | Overexpression of the Bacteriophage T4 <i>motB</i> Gene Alters H-NS Dependent Repression of Specific Host DNA |
| title_short | Overexpression of the Bacteriophage T4 <i>motB</i> Gene Alters H-NS Dependent Repression of Specific Host DNA |
| title_sort | overexpression of the bacteriophage t4 i motb i gene alters h ns dependent repression of specific host dna |
| topic | bacteriophage T4 MotB H-NS nucleoid host takeover DNA-binding protein |
| url | https://www.mdpi.com/1999-4915/13/1/84 |
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