PMID: 9192626Jun 24, 1997Paper

DNA damage-dependent transcriptional arrest and termination of RNA polymerase II elongation complexes in DNA template containing HIV-1 promoter

Proceedings of the National Academy of Sciences of the United States of America
Z Wang, T M Rana

Abstract

We have developed a new biochemical method to isolate a homogeneous population of RNA polymerase II (RNA pol II) elongation complexes arrested at a DNA damage site. The method involves triple-helix formation at a predetermined site in DNA template with a third strand labeled with psoralen at its 5'-end and a biotin at the 3'-end. After triplex formation and near-ultraviolet irradiation (360 nm), DNA templates modified with psoralen were immobilized on streptavidin-coated magnetic beads and used for in vitro transcription reactions with HeLa nuclear extracts. Separation of magnetic beads from solution results in isolation of arrested elongation complexes on the immobilized DNA templates. We have applied the method to arrest RNA pol II elongation complexes on a DNA template containing HIV-1 promoter. Our results indicate that psoralen crosslink in the template strand efficiently arrests elongation complexes, and psoralen monoadducts terminate transcription. Our results also demonstrate that a triple-helical structure stabilized by an intercalator, acridine, attached to the third strand of the helix inhibits transcription by a termination pathway. Isolation of stable RNA pol II elongation complexes arrested at DNA damage sites is ...Continue Reading

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Citations

Sep 1, 2001·The Journal of Gene Medicine·M Faria, C Giovannangeli
May 21, 2005·Mutation Research·Silvia Tornaletti
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Apr 13, 2000·Proceedings of the National Academy of Sciences of the United States of America·M FariaC Giovannangeli
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Mar 21, 2003·The Journal of Biological Chemistry·Virginia S KalogerakiPhilip C Hanawalt
Mar 17, 2007·The Journal of Biological Chemistry·Takayuki NojimaMasatoshi Hagiwara

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