Genetic Requirements for Sensitivity of Bacteriophage T7 to Dideoxythymidine
| العنوان: | Genetic Requirements for Sensitivity of Bacteriophage T7 to Dideoxythymidine |
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| المؤلفون: | Ngoc Quang Tran, Stanley Tabor, Charles C. Richardson |
| المصدر: | Journal of Bacteriology. 196:2842-2850 |
| بيانات النشر: | American Society for Microbiology, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | DNA-Directed DNA Polymerase, medicine.disease_cause, Thymidine Kinase, Microbiology, Bacteriophage, Gene Knockout Techniques, Viral Proteins, chemistry.chemical_compound, Bacteriophage T7, parasitic diseases, Escherichia coli, medicine, Thymine Nucleotides, Phosphorylation, Thymidine phosphorylase, Molecular Biology, Nucleic Acid Synthesis Inhibitors, Sequence Deletion, biology, Escherichia coli Proteins, Pyrimidine Phosphorylases, T7 DNA polymerase, Articles, biology.organism_classification, Molecular biology, Thymine, chemistry, Biochemistry, Thymidine kinase, DNA, Viral, Thymidine, DNA, Dideoxynucleotides |
| الوصف: | We previously reported that the presence of dideoxythymidine (ddT) in the growth medium selectively inhibits the ability of bacteriophage T7 to infect Escherichia coli by inhibiting phage DNA synthese (N. Q. Tran, L. F. Rezende, U. Qimron, C. C. Richardson, and S. Tabor, Proc. Natl. Acad. Sci. U. S. A. 105:9373–9378, 2008, doi:10.1073/pnas.0804164105). In the presence of T7 gene 1.7 protein, ddT is taken up into the E. coli cell and converted to ddTTP. ddTTP is incorporated into DNA as ddTMP by the T7 DNA polymerase, resulting in chain termination. We have identified the pathway by which exogenous ddT is converted to ddTTP. The pathway consists of ddT transport by host nucleoside permeases and phosphorylation to ddTMP by the host thymidine kinase. T7 gene 1.7 protein phosphorylates ddTMP and ddTDP, resulting in ddTTP. A 74-residue peptide of the gene 1.7 protein confers ddT sensitivity to the same extent as the 196-residue wild-type gene 1.7 protein. We also show that cleavage of thymidine to thymine and deoxyribose-1-phosphate by the host thymidine phosphorylase greatly increases the sensitivity of phage T7 to ddT. Finally, a mutation in T7 DNA polymerase that leads to discrimination against the incorporation of ddTMP eliminates ddT sensitivity. |
| تدمد: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.01718-14 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4f0d500aaca5be0bbded854313702d5b https://doi.org/10.1128/jb.01718-14 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....4f0d500aaca5be0bbded854313702d5b |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10985530 00219193 |
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| DOI: | 10.1128/jb.01718-14 |