NDLI: Potent stimulation of transcription-coupled DNA supercoilingby sequence-specific DNA-binding proteins

Content Provider	PubMed Central
Author	Leng, Fenfei Roger, Mcmacken
Copyright Year	2002
Abstract	Transcription by RNA polymerase can stimulate localized DNAsupercoiling in Escherichia coli. Invivo, there is extensive experimental support for a“twin-domain” model in which positive DNA supercoils aregenerated ahead of a translocating RNA polymerase complex and negativesupercoils are formed behind it. Negative supercoils accumulatein the template DNA because the positive supercoils are preferentiallyremoved by cellular topoisomerase action. Yet, in vitro,clear and convincing support for the twin-domain mechanism has beenlacking. In this article, we reconcile this inconsistency by showingthat, in a defined in vitro system with plasmid DNAtemplates, a variety of sequence-specific DNA-binding proteins, such asthe bacteriophage λ O replication initiator or the E.coli lactose or galactose repressors, strikingly stimulatetranscription-coupled DNA supercoiling. We demonstrate further thatthis stimulation requires the presence in the DNA template of arecognition sequence for the relevant DNA-binding protein and dependson the production of long RNA chains by an RNA polymerase. Our data aremost consistent with a model in which specific DNA-binding proteinsfacilitate a twin-domain mechanism to enhance DNA supercoiling duringtranscription. More precisely, we suggest that some nucleoproteincomplexes, perhaps those that contain sharply bent DNA, can formbarriers that impede the diffusion and merger of independentchromosomal supercoil domains. Localization of DNA supercoilsby nucleoprotein complexes may serve as a generalmechanism for modulating DNA transactions that are sensitive toDNA superhelicity.
Related Links	http://dx.doi.org/10.1073/pnas.142002099
Ending Page	9144
Page Count	6
Starting Page	9139
File Format	PDF
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	14
Volume Number	99
Language	English
Publisher	National Academy of Sciences
Publisher Date	2002-07-09
Access Restriction	Open
Rights Holder	National Academy of Sciences
Subject Keyword	General Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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