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Harden, Timothy T. ; Herlambang, Karina S. ; Chamberlain, Mathew ; Lalanne, Jean-Benoît ; Wells, Christopher D. ; Li, Gene-Wei ; Landick, Robert ; Hochschild, Ann ; Kondev, Jane ; Gelles, Jeff ( , Nature Communications)
Abstract RNA polymerases (RNAPs) transcribe genes through a cycle of recruitment to promoter DNA, initiation, elongation, and termination. After termination, RNAP is thought to initiate the next round of transcription by detaching from DNA and rebinding a new promoter. Here we use single-molecule fluorescence microscopy to observe individual RNAP molecules after transcript release at a terminator. Following termination, RNAP almost always remains bound to DNA and sometimes exhibits one-dimensional sliding over thousands of basepairs. Unexpectedly, the DNA-bound RNAP often restarts transcription, usually in reverse direction, thus producing an antisense transcript. Furthermore, we report evidence of this secondary initiation in live cells, using genome-wide RNA sequencing. These findings reveal an alternative transcription cycle that allows RNAP to reinitiate without dissociating from DNA, which is likely to have important implications for gene regulation.
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Vincent, Ben J. ; Staller, Max V. ; Lopez-Rivera, Francheska ; Bragdon, Meghan D. ; Pym, Edward C. ; Biette, Kelly M. ; Wunderlich, Zeba ; Harden, Timothy T. ; Estrada, Javier ; DePace, Angela H. ; et al ( , PLOS Genetics)
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Harden, Timothy T. ; Wells, Christopher D. ; Friedman, Larry J. ; Landick, Robert ; Hochschild, Ann ; Kondev, Jane ; Gelles, Jeff ( , Proceedings of the National Academy of Sciences)