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• W2145785862 abstract "Coliphage N4 virion RNA polymerase (vRNAP), the most distantly related member of the T7-like family of RNA polymerases, is responsible for transcription of the early genes of the linear double-stranded DNA phage genome. Escherichia coli single-stranded DNA-binding protein ( Eco SSB) is required for N4 early transcription in vivo , as well as for in vitro transcription on super-coiled DNA templates containing vRNAP promoters. In contrast to other DNA-dependent RNA polymerases, vRNAP initiates transcription on single-stranded, promoter-containing templates with in vivo specificity; however, the RNA product is not displaced, thus limiting template usage to one round. We show that Eco SSB activates vRNAP transcription at limiting single-stranded template concentrations through template recycling. Eco SSB binds to the template and to the nascent transcript and prevents the formation of a transcriptionally inert RNA:DNA hybrid. Using C-terminally truncated Eco SSB mutant proteins, human mitochondrial SSB ( Hs mt SSB), phage P1 SSB, and F episome-encoded SSB, as well as a Hs mt- Eco SSB chimera, we have mapped a determinant of template recycling to the C-terminal amino acids of Eco SSB. T7 RNAP contains an amino-terminal domain responsible for binding the RNA product as it exits from the enzyme. No sequence similarity to this domain exists in vRNAP. Hereby, we propose a unique role for Eco SSB: It functionally substitutes in N4 vRNAP for the N-terminal domain of T7 RNAP responsible for RNA binding." @default.
• W2145785862 created "2016-06-24" @default.
• W2145785862 creator A5018195491 @default.
• W2145785862 creator A5021415693 @default.
• W2145785862 date "2003-07-22" @default.
• W2145785862 modified "2023-10-16" @default.
• W2145785862 title "Escherichia coli single-stranded DNA-binding protein mediates template recycling during transcription by bacteriophage N4 virion RNA polymerase" @default.
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• W2145785862 doi "https://doi.org/10.1073/pnas.1133325100" @default.
• W2145785862 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/170904" @default.
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• W2145785862 hasPublicationYear "2003" @default.
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