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    Cluster K Mycobacteriophages: Insights into the Evolutionary Origins of Mycobacteriophage TM4

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    Five newly isolated mycobacteriophages –Angelica, CrimD, Adephagia, Anaya, and Pixie – have similar genomic architectures to mycobacteriophage TM4, a previously characterized phage that is widely used in mycobacterial genetics. The nucleotide sequence similarities warrant grouping these into Cluster K, with subdivision into three subclusters: K1, K2, and K3. Although the overall genome architectures of these phages are similar, TM4 appears to have lost at least two segments of its genome, a central region containing the integration apparatus, and a segment at the right end. This suggests that TM4 is a recent derivative of a temperate parent, resolving a long-standing conundrum about its biology, in that it was reportedly recovered from a lysogenic strain of Mycobacterium avium, but it is not capable of forming lysogens in any mycobacterial host. Like TM4, all of the Cluster K phages infect both fast- and slow-growing mycobacteria, and all of them – with the exception of TM4 – form stable lysogens in both Mycobacterium smegmatis and Mycobacterium tuberculosis; immunity assays show that all five of these phages share the same immune specificity. TM4 infects these lysogens suggesting that it was either derived from a heteroimmune temperate parent or that it has acquired a virulent phenotype. We have also characterized a widely-used conditionally replicating derivative of TM4 and identified mutations conferring the temperature-sensitive phenotype. All of the Cluster K phages contain a series of well conserved 13 bp repeats associated with the translation initiation sites of a subset of the genes; approximately one half of these contain an additional sequence feature composed of imperfectly conserved 17 bp inverted repeats separated by a variable spacer. The K1 phages integrate into the host tmRNA and the Cluster K phages represent potential new tools for the genetics of M. tuberculosis and related species

    Cluster k mycobacteriophages: Insights into the evolutionary origins of mycobacteriophage tm4

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    Five newly isolated mycobacteriophages -Angelica, CrimD, Adephagia, Anaya, and Pixie - have similar genomic architectures to mycobacteriophage TM4, a previously characterized phage that is widely used in mycobacterial genetics. The nucleotide sequence similarities warrant grouping these into Cluster K, with subdivision into three subclusters: K1, K2, and K3. Although the overall genome architectures of these phages are similar, TM4 appears to have lost at least two segments of its genome, a central region containing the integration apparatus, and a segment at the right end. This suggests that TM4 is a recent derivative of a temperate parent, resolving a long-standing conundrum about its biology, in that it was reportedly recovered from a lysogenic strain of Mycobacterium avium, but it is not capable of forming lysogens in any mycobacterial host. Like TM4, all of the Cluster K phages infect both fast- and slow-growing mycobacteria, and all of them - with the exception of TM4 - form stable lysogens in both Mycobacterium smegmatis and Mycobacterium tuberculosis; immunity assays show that all five of these phages share the same immune specificity. TM4 infects these lysogens suggesting that it was either derived from a heteroimmune temperate parent or that it has acquired a virulent phenotype. We have also characterized a widely-used conditionally replicating derivative of TM4 and identified mutations conferring the temperature-sensitive phenotype. All of the Cluster K phages contain a series of well conserved 13 bp repeats associated with the translation initiation sites of a subset of the genes; approximately one half of these contain an additional sequence feature composed of imperfectly conserved 17 bp inverted repeats separated by a variable spacer. The K1 phages integrate into the host tmRNA and the Cluster K phages represent potential new tools for the genetics of M. tuberculosis and related species. © 2011 Pope et al.Fil: Pope, Welkin H.. University of Pittsburgh; Estados UnidosFil: Ferreira, Christina M.. University of Pittsburgh; Estados UnidosFil: Jacobs Sera, Deborah. University of Pittsburgh; Estados UnidosFil: Benjamin, Robert C.. University of North Texas; Estados UnidosFil: Davis, Ariangela J.. Calvin College; Estados UnidosFil: DeJong, Randall J.. Calvin College; Estados UnidosFil: Elgin, Sarah C. R.. Washington University in St. Louis; Estados UnidosFil: Guilfoile, Forrest R.. University of Pittsburgh; Estados UnidosFil: Forsyth, Mark H.. The College Of William And Mary; Estados UnidosFil: Harris, Alexander D.. Calvin College; Estados UnidosFil: Harvey, Samuel E.. The College Of William And Mary; Estados UnidosFil: Hughes, Lee E.. University of North Texas; Estados UnidosFil: Hynes, Peter M.. Washington University in St. Louis; Estados UnidosFil: Jackson, Arrykka S.. The College Of William And Mary; Estados UnidosFil: Jalal, Marilyn D.. University of North Texas; Estados UnidosFil: MacMurray, Elizabeth A.. The College Of William And Mary; Estados UnidosFil: Manley, Coreen M.. University of North Texas; Estados UnidosFil: McDonough, Molly J.. The College Of William And Mary; Estados UnidosFil: Mosier, Jordan L.. University of North Texas; Estados UnidosFil: Osterbann, Larissa J.. Calvin College; Estados UnidosFil: Rabinowitz, Hannah S.. Washington University in St. Louis; Estados UnidosFil: Rhyan, Corwin N.. Washington University in St. Louis; Estados UnidosFil: Russell, Daniel A.. University of Pittsburgh; Estados UnidosFil: Saha, Margaret S.. The College Of William And Mary; Estados UnidosFil: Shaffer, Christopher D.. Washington University in St. Louis; Estados UnidosFil: Simon, Stephanie E.. University of North Texas; Estados UnidosFil: Sims, Erika F.. Washington University in St. Louis; Estados UnidosFil: Tovar, Isabel G.. University of North Texas; Estados UnidosFil: Weisser, Emilie G.. Washington University in St. Louis; Estados UnidosFil: Wertz, John T.. Calvin College; Estados UnidosFil: Weston-Hafer, Kathleen A.. Washington University in St. Louis; Estados UnidosFil: Williamson, Kurt E.. The College Of William And Mary; Estados UnidosFil: Zhang, Bo. Washington University in St. Louis; Estados UnidosFil: Cresawn, Steven G.. James Madison University; Estados UnidosFil: Jain, Paras. Albert Einstein College Of Medicine Of Yeshiva University; Estados UnidosFil: Piuri, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Pittsburgh; Estados UnidosFil: Jacobs, William R.. Albert Einstein College Of Medicine Of Yeshiva University; Estados UnidosFil: Hendrix, Roger W.. University of Pittsburgh; Estados UnidosFil: Hatfull, Graham F.. University of Pittsburgh; Estados Unido
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