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Purification and in vitro DNA-binding specificity of the Bacillus subtilis phage Φ105 repressor

Luc Van Kaer, Marc Van Montagu UGent and Patrick Dhaese UGent (1989) JOURNAL OF BIOLOGICAL CHEMISTRY. 264(25). p.14784-14791
abstract
The Bacillus subtilis phage phi 105 repressor, a lambda repressor-like transcriptional regulatory protein, was overproduced in Escherichia coli and purified to near homogeneity in order to examine its in vitro DNA-binding properties. The active form of repressor appears to be a tetramer. DNase I protection experiments demonstrate that repressor can specifically bind to six distinct sites, all located within the phi 105 EcoRI-F immunity region (immF). Three of these sites had been identified earlier as functional operators by genetic analysis. They share a common 14-base pair, asymmetric "core" sequence, 5'-GACGGAAATACAAG-3', termed OR. The three additional sites show significant homology with OR. For an individual binding site, hydroxyl-radical footprinting reveals symmetrical repressor-DNA interactions established via one side of the helix. Dimethyl sulfate protection indicates that guanines at the conserved OR base pair positions 1, 3, and 4 may participate in sequence-specific interactions with repressor in agreement with a previously proposed recognition model. However, since the OR sequence is not symmetrical with respect to this GNCG motif, at present it remains difficult to completely understand the molecular basis of this interaction.
Please use this url to cite or link to this publication:
author
organization
alternative title
Purification and invitro DNA-binding specificity of the Bacillus subtilis phage phi-105 repressor
year
type
journalArticle (original)
publication status
published
subject
journal title
JOURNAL OF BIOLOGICAL CHEMISTRY
J. Biol. Chem.
volume
264
issue
25
pages
14784 - 14791
Web of Science type
Article
ISSN
0021-9258
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1910400
handle
http://hdl.handle.net/1854/LU-1910400
alternative location
http://www.jbc.org/content/264/25/14784.abstract
date created
2011-09-28 11:56:36
date last changed
2016-12-19 15:46:25
@article{1910400,
  abstract     = {The Bacillus subtilis phage phi 105 repressor, a lambda repressor-like transcriptional regulatory protein, was overproduced in Escherichia coli and purified to near homogeneity in order to examine its in vitro DNA-binding properties. The active form of repressor appears to be a tetramer. DNase I protection experiments demonstrate that repressor can specifically bind to six distinct sites, all located within the phi 105 EcoRI-F immunity region (immF). Three of these sites had been identified earlier as functional operators by genetic analysis. They share a common 14-base pair, asymmetric {\textacutedbl}core{\textacutedbl} sequence, 5'-GACGGAAATACAAG-3', termed OR. The three additional sites show significant homology with OR. For an individual binding site, hydroxyl-radical footprinting reveals symmetrical repressor-DNA interactions established via one side of the helix. Dimethyl sulfate protection indicates that guanines at the conserved OR base pair positions 1, 3, and 4 may participate in sequence-specific interactions with repressor in agreement with a previously proposed recognition model. However, since the OR sequence is not symmetrical with respect to this GNCG motif, at present it remains difficult to completely understand the molecular basis of this interaction.},
  author       = {Van Kaer, Luc and Van Montagu, Marc and Dhaese, Patrick},
  issn         = {0021-9258},
  journal      = {JOURNAL OF BIOLOGICAL CHEMISTRY},
  language     = {eng},
  number       = {25},
  pages        = {14784--14791},
  title        = {Purification and in vitro DNA-binding specificity of the Bacillus subtilis phage \ensuremath{\Phi}105 repressor},
  url          = {http://www.jbc.org/content/264/25/14784.abstract},
  volume       = {264},
  year         = {1989},
}

Chicago
Van Kaer, Luc, Marc Van Montagu, and Patrick Dhaese. 1989. “Purification and in Vitro DNA-binding Specificity of the Bacillus Subtilis Phage Φ105 Repressor.” Journal of Biological Chemistry 264 (25): 14784–14791.
APA
Van Kaer, L., Van Montagu, M., & Dhaese, P. (1989). Purification and in vitro DNA-binding specificity of the Bacillus subtilis phage Φ105 repressor. JOURNAL OF BIOLOGICAL CHEMISTRY, 264(25), 14784–14791.
Vancouver
1.
Van Kaer L, Van Montagu M, Dhaese P. Purification and in vitro DNA-binding specificity of the Bacillus subtilis phage Φ105 repressor. JOURNAL OF BIOLOGICAL CHEMISTRY. 1989;264(25):14784–91.
MLA
Van Kaer, Luc, Marc Van Montagu, and Patrick Dhaese. “Purification and in Vitro DNA-binding Specificity of the Bacillus Subtilis Phage Φ105 Repressor.” JOURNAL OF BIOLOGICAL CHEMISTRY 264.25 (1989): 14784–14791. Print.