"Antiparallel" DNA loop in Gal repressosome visualized by atomic force microscopy

Konstantin Virnik, Yuri L. Lyubchenko, Mikhail A. Karymov, Paul Dahlgren, Michael Y. Tolstorukov, Szabolcs Semsey, Victor B. Zhurkin, Sankar Adhya

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

DNA looping is often involved in positive and negative regulation of gene transcription in both prokaryotes and eukaryotes. The transcription of the gal operon of Escherichia coli from two overlapping promoters P1 and P2 is negatively regulated via Gal repressosome assembly. It involves binding of two dimeric Gal repressor proteins (GalR) to two operators, OE and O I, flanking the two promoters, and formation of 113 bp DNA loop due to tetramerization of the two bound GalR dimers. The process requires negatively supercoiled DNA and the presence of the histone-like protein HU. Previous modeling of the repressosome based on evaluation of DNA elastic energy suggested a mutual antiparallel, rather than parallel, orientation of the two gal operators in an under-twisted DNA loop. To visualize the Gal loop by atomic force microscopy (AFM), plasmid DNA molecules were constructed with increased distance between the two operators. The AFM results demonstrated the formation of an antiparallel DNA loop in the Gal repressosome consistent with our earlier hypothesis. Importantly, the overall shape of the GalR mediated loop proved to be indistinguishable from that in the chimerical loop of the same size containing two lac operators (instead of two gal operators) and formed by LacI. In addition, a possibility of the gal operon repression mediated by GalR in the absence of HU was shown in the new DNA constructs. Implications of these findings for the DNA structural organization in bacterial nucleoid are discussed.

Original languageEnglish (US)
Pages (from-to)53-63
Number of pages11
JournalJournal of Molecular Biology
Volume334
Issue number1
DOIs
StatePublished - Nov 14 2003

Fingerprint

Atomic Force Microscopy
DNA
Operon
Superhelical DNA
Eukaryota
Histones
Plasmids
Escherichia coli
Galactose repressor proteins

Keywords

  • AFM
  • DNA loop
  • Minicircles
  • Repression
  • Transcription

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Virnik, K., Lyubchenko, Y. L., Karymov, M. A., Dahlgren, P., Tolstorukov, M. Y., Semsey, S., ... Adhya, S. (2003). "Antiparallel" DNA loop in Gal repressosome visualized by atomic force microscopy. Journal of Molecular Biology, 334(1), 53-63. https://doi.org/10.1016/j.jmb.2003.09.030

"Antiparallel" DNA loop in Gal repressosome visualized by atomic force microscopy. / Virnik, Konstantin; Lyubchenko, Yuri L.; Karymov, Mikhail A.; Dahlgren, Paul; Tolstorukov, Michael Y.; Semsey, Szabolcs; Zhurkin, Victor B.; Adhya, Sankar.

In: Journal of Molecular Biology, Vol. 334, No. 1, 14.11.2003, p. 53-63.

Research output: Contribution to journalArticle

Virnik, K, Lyubchenko, YL, Karymov, MA, Dahlgren, P, Tolstorukov, MY, Semsey, S, Zhurkin, VB & Adhya, S 2003, '"Antiparallel" DNA loop in Gal repressosome visualized by atomic force microscopy', Journal of Molecular Biology, vol. 334, no. 1, pp. 53-63. https://doi.org/10.1016/j.jmb.2003.09.030
Virnik, Konstantin ; Lyubchenko, Yuri L. ; Karymov, Mikhail A. ; Dahlgren, Paul ; Tolstorukov, Michael Y. ; Semsey, Szabolcs ; Zhurkin, Victor B. ; Adhya, Sankar. / "Antiparallel" DNA loop in Gal repressosome visualized by atomic force microscopy. In: Journal of Molecular Biology. 2003 ; Vol. 334, No. 1. pp. 53-63.
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