{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T23:20:44Z","timestamp":1789687244121,"version":"4.0.1"},"reference-count":45,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2018,6,26]],"date-time":"2018-06-26T00:00:00Z","timestamp":1529971200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The lytic bacteriophage T4 employs multiple phage-encoded early proteins to takeover the Escherichia coli host. However, the functions of many of these proteins are not known. In this study, we have characterized the T4 early gene motB, located in a dispensable region of the T4 genome. We show that heterologous production of MotB is highly toxic to E. coli, resulting in cell death or growth arrest depending on the strain and that the presence of motB increases T4 burst size 2-fold. Previous work suggested that motB affects middle gene expression, but our transcriptome analyses of T4 motBam vs. T4 wt infections reveal that only a few late genes are mildly impaired at 5 min post-infection, and expression of early and middle genes is unaffected. We find that MotB is a DNA-binding protein that binds both unmodified host and T4 modified [(glucosylated, hydroxymethylated-5 cytosine, (GHme-C)] DNA with no detectable sequence specificity. Interestingly, MotB copurifies with the host histone-like proteins, H-NS and StpA, either directly or through cobinding to DNA. We show that H-NS also binds modified T4 DNA and speculate that MotB may alter how H-NS interacts with T4 DNA, host DNA, or both, thereby improving the growth of the phage.<\/jats:p>","DOI":"10.3390\/v10070343","type":"journal-article","created":{"date-parts":[[2018,6,26]],"date-time":"2018-06-26T10:40:50Z","timestamp":1530009650000},"page":"343","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["The Bacteriophage T4 MotB Protein, a DNA-Binding Protein, Improves Phage Fitness"],"prefix":"10.3390","volume":"10","author":[{"given":"Jennifer","family":"Patterson-West","sequence":"first","affiliation":[{"name":"Gene 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 20892-0830, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa","family":"Arroyo-Mendoza","sequence":"additional","affiliation":[{"name":"Gene 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 20892-0830, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng-Lun","family":"Hsieh","sequence":"additional","affiliation":[{"name":"Gene 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 20892-0830, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danielle","family":"Harrison","sequence":"additional","affiliation":[{"name":"Gene 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 20892-0830, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morgan M.","family":"Walker","sequence":"additional","affiliation":[{"name":"Gene 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 20892-0830, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leslie","family":"Knipling","sequence":"additional","affiliation":[{"name":"Gene 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 20892-0830, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah M.","family":"Hinton","sequence":"additional","affiliation":[{"name":"Gene 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 20892-0830, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,6,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2153","DOI":"10.1007\/s11274-014-1655-7","article-title":"Natural solution to antibiotic resistance: Bacteriophages, \u201cthe living drugs\u201c","volume":"30","author":"Jassim","year":"2014","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"699","DOI":"10.2174\/138920312804871193","article-title":"Learning from bacteriophages\u2014Advantages and limitations of phage and phage-encoded protein applications","volume":"13","author":"Maciejewska","year":"2012","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1016\/j.coviro.2011.06.009","article-title":"Bacteriophages and their genomes","volume":"1","author":"Hatfull","year":"2011","journal-title":"Curr. Opin. Virol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1128\/MMBR.67.1.86-156.2003","article-title":"Bacteriophage T4 genome","volume":"67","author":"Miller","year":"2003","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1186\/1743-422X-7-289","article-title":"Transcriptional control in the prereplicative phase of T4 development","volume":"7","author":"Hinton","year":"2010","journal-title":"Virol. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7974","DOI":"10.1093\/nar\/gkw656","article-title":"Visualizing the phage T4 activated transcription complex of DNA and E. coli RNA polymerase","volume":"44","author":"James","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1186\/1743-422X-7-288","article-title":"Transcription of the T4 late genes","volume":"7","author":"Geiduschek","year":"2010","journal-title":"Virol. J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1042\/BST20160190","article-title":"Structure and function of bacterial H-NS protein","volume":"44","author":"Grainger","year":"2016","journal-title":"Biochem. Soc. Trans."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Navarre, W.W. (2010). H-NS as a defence system. Bacterial Chromatin, Springer.","DOI":"10.1007\/978-90-481-3473-1_13"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"27046","DOI":"10.1074\/jbc.M114.590851","article-title":"The T4 phage DNA mimic protein Arn inhibits the DNA binding activity of the bacterial histone-like protein H-NS","volume":"289","author":"Ho","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1128\/jb.149.2.694-699.1982","article-title":"Physical mapping and cloning of bacteriophage T4 anti-restriction endonuclease gene","volume":"149","author":"Dharmalingam","year":"1982","journal-title":"J. Bacteriol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4881","DOI":"10.1128\/JB.05198-11","article-title":"The 5.5 protein of phage T7 inhibits H-NS through interactions with the central oligomerization domain","volume":"193","author":"Ali","year":"2011","journal-title":"J. Bacteriol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1242","DOI":"10.3389\/fmicb.2015.01242","article-title":"Antibacterial phage orfans of pseudomonas aeruginosa phage LUZ24 reveal a novel MvaT inhibiting protein","volume":"6","author":"Wagemans","year":"2015","journal-title":"Front. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1111\/j.1365-2958.1996.tb02565.x","article-title":"Integration host factor alleviates the H-NS-mediated repression of the early promoter of bacteriophage Mu","volume":"21","author":"Hillebrand","year":"1996","journal-title":"Mol. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/0378-1119(84)90103-3","article-title":"Regulation of MU transposition. I. Localization of the presumed recognition sites for HIMD and NER functions controlling bacteriophage MU transcription","volume":"30","author":"Goosen","year":"1984","journal-title":"Gene"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/0022-2836(85)90343-2","article-title":"New control elements of bacteriophage-T4 pre-replicative transcription","volume":"182","author":"Pulitzer","year":"1985","journal-title":"J. Mol. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/S0378-1119(98)00238-8","article-title":"Several new bacteriophage T4 genes, mapped by sequencing deletion endpoints between genes 56 (DCTPase) and DDA (a DNA-dependent atpase-helicase) modulate transcription","volume":"223","author":"Mosig","year":"1998","journal-title":"Gene"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1016\/S0021-9258(18)89779-4","article-title":"The T4 Mot protein functions as part of a pre-replicative DNA-protein complex","volume":"260","author":"Uzan","year":"1985","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1111\/j.1365-2958.2008.06276.x","article-title":"A basic\/hydrophobic cleft of the T4 activator MotA interacts with the C-terminus of E. coli sigma(70) to activate middle gene transcription","volume":"69","author":"Bonocora","year":"2008","journal-title":"Mol. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/0076-6879(90)85008-C","article-title":"Use of T7 RNA-polymerase to direct expression of cloned genes","volume":"185","author":"Studier","year":"1990","journal-title":"Methods Enzymol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1007\/BF00332996","article-title":"The isolation and characterization of TabR bacteria\u2014Hosts that restrict bacteriophage-T4 rII mutants","volume":"188","author":"Nelson","year":"1982","journal-title":"Mol. Gen. Genet."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/0022-2836(78)90058-X","article-title":"Isolation and characterization of conditional lethal mutations in the mot gene of bacteriophage-T4","volume":"126","author":"Mattson","year":"1978","journal-title":"J. Mol. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1126\/science.322279","article-title":"Screening lambdagt recombinant clones by hybridization to single plaques in situ","volume":"196","author":"Benton","year":"1977","journal-title":"Science"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6133","DOI":"10.1128\/jb.178.21.6133-6139.1996","article-title":"An N-terminal mutation in the bacteriophage T4 motA gene yields a protein that binds DNA but is defective for activation of transcription","volume":"178","author":"Gerber","year":"1996","journal-title":"J. Bacteriol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4391","DOI":"10.1128\/JB.181.14.4391-4396.1999","article-title":"Lysis and lysis inhibition in bacteriophage T4: rV mutations reside in the holin T gene","volume":"181","author":"Dressman","year":"1999","journal-title":"J. Bacteriol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Karam, J.D., Kreuzer, K.N., Mosig, G., Hall, D.H., Eiserling, F.A., Black, L.W., Spicer, E.K., Kutter, E., Carlson, K., and Miller, E.S. (1994). Single-step growth. Molecular Biology of Bacteriophage T4, American Society of Microbiology.","DOI":"10.1128\/ecosalplus.bacteriophage-t4"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"14432","DOI":"10.1016\/S0021-9258(18)71697-9","article-title":"Transcript analyses of the uvsX-40-41 region of bacteriophage T4. Changes in the RNA as infection proceeds","volume":"264","author":"Hinton","year":"1989","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Schroeder, A., Mueller, O., Stocker, S., Salowsky, R., Leiber, M., Gassmann, M., Lightfoot, S., Menzel, W., Granzow, M., and Ragg, T. (2006). The RIN: An RNA integrity number for assigning integrity values to RNA measurements. BMC Mol. Biol., 7.","DOI":"10.1186\/1471-2199-7-3"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Chevallereau, A., Blasdel, B.G., De Smet, J., Monot, M., Zimmermann, M., Kogadeeva, M., Sauer, U., Jorth, P., Whiteley, M., and Debarbieux, L. (2016). Next-generation \u201c-omics\u201d approaches reveal a massive alteration of host RNA metabolism during bacteriophage infection of Pseudomonas aeruginosa. PLoS Genet., 12.","DOI":"10.1371\/journal.pgen.1006134"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Leskinen, K., Blasdel, B.G., Lavigne, R., and Skurnik, M. (2016). Rna-sequencing reveals the progression of phage-host interactions between \u03d5R1-37 and Yersinia enterocolitica. Viruses, 8.","DOI":"10.3390\/v8040111"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1158\/1535-7163.MCT-07-0573","article-title":"MicroRNAs modulate the chemosensitivity of tumor cells","volume":"7","author":"Blower","year":"2008","journal-title":"Mol. Cancer Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1289","DOI":"10.1093\/bioinformatics\/btm091","article-title":"Enhancements and modifications of primer design program Primer3","volume":"23","author":"Koressaar","year":"2007","journal-title":"Bioinformatics"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e115","DOI":"10.1093\/nar\/gks596","article-title":"Primer3\u2014New capabilities and interfaces","volume":"40","author":"Untergasser","year":"2012","journal-title":"Nucleic Acids Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/0022-2836(88)90311-7","article-title":"Transcriptional activation of bacteriophage-T4 middle promoters by the MotA protein","volume":"199","author":"Guild","year":"1988","journal-title":"J. Mol. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"5580","DOI":"10.1128\/JB.00338-10","article-title":"A mutation within the \u03b2 subunit of Escherichia coli RNA polymerase impairs transcription from bacteriophage T4 middle promoters","volume":"192","author":"James","year":"2010","journal-title":"J. Bacteriol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"18034","DOI":"10.1016\/S0021-9258(18)55233-9","article-title":"Transcription from a bacteriophage-T4 middle promoter using T4 motA protein and phage-modified RNA-polymerase","volume":"266","author":"Hinton","year":"1991","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"R37","DOI":"10.1186\/gb-2012-13-5-r37","article-title":"Comparative multi-omics systems analysis of Escherichia coli strains B and K-12","volume":"13","author":"Yoon","year":"2012","journal-title":"Genome Biol."},{"key":"ref_38","first-page":"62","article-title":"Validity of nucleic acid purities monitored by 260 nm\/280 nm absorbance ratios","volume":"18","author":"Glasel","year":"1995","journal-title":"Biotechniques"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5321","DOI":"10.1093\/nar\/12.13.5321","article-title":"H1A, an E. coli DNA-binding protein which accumulates in stationary phase, strongly compacts DNA in vitro","volume":"12","author":"Spassky","year":"1984","journal-title":"Nucleic Acids Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1038\/nsmb1233","article-title":"H-Ns cooperative binding to high-affinity sites in a regulatory element results in transcriptional silencing","volume":"14","author":"Bouffartigues","year":"2007","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"9593","DOI":"10.1039\/c3sm51214b","article-title":"Effect of H-NS on the elongation and compaction of single DNA molecules in a nanospace","volume":"9","author":"Zhang","year":"2013","journal-title":"Soft Matter"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Karam, J.D. (1994). Regulation of middle-mode transcription. Molecular Biology of Bacteriophage T4, American Society of Microbiology.","DOI":"10.1128\/ecosalplus.bacteriophage-t4"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/0378-1119(90)90053-T","article-title":"The bacteriophage T4 gene MRH whose product inhibits late T4 gene expression in an Escherichia coli rpoh (sigma 32) mutant","volume":"88","author":"Frazier","year":"1990","journal-title":"Gene"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Karam, J. (1994). Gel analysis of T4 prereplicative proteins. Molecular Biology of Bacteriophage T4, American Society of Microbiology.","DOI":"10.1128\/ecosalplus.bacteriophage-t4"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"700","DOI":"10.1128\/jvi.5.6.700-708.1970","article-title":"Isolation of bacteriophage T4 mutants defective in the ability to degrade host deoxyribonucleic acid","volume":"5","author":"Warner","year":"1970","journal-title":"J. Virol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/10\/7\/343\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:10:14Z","timestamp":1760195414000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/10\/7\/343"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,6,26]]},"references-count":45,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2018,7]]}},"alternative-id":["v10070343"],"URL":"https:\/\/doi.org\/10.3390\/v10070343","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,6,26]]}}}