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Flow linear dichroism and electron microscopic analysis of protein-DNA complexes of a mutant UvrB protein that binds to but cannot kink DNA.

Publication ,  Journal Article
Hsu, DS; Takahashi, M; Delagoutte, E; Bertrand-Burggraf, E; Wang, YH; Norden, B; Fuchs, RP; Griffith, J; Sancar, A
Published in: J Mol Biol
September 2, 1994

(A)BC excinuclease of Escherichia coli is the enzymatic activity resulting from sequential and partially overlapping actions of UvrA, UvrB, and UvrC protein. UvrA is a molecular matchmaker which promotes the formation of a stable UvrB-damaged DNA complex in which the DNA is kinked by about 130 degrees. The UvrB-DNA complex is then recognized by UvrC and two incisions are made in the DNA by the joint actions of UvrC and UvrB. A mutant of UvrB (D478A) can be loaded onto the DNA but it does not interact with UvrC to cause a nick 3' to the lesion. Based on the lack of a DNase-I-hypersensitive site in the footprint of the mutant, it was proposed that the lack of incision was due to the inability of the mutant UvrB to kink the DNA. In the current study we have investigated the interaction of the mutant UvrB with DNA using two biophysical methods, flow linear dichroism and electron microscopy. Both methods reveal that the mutant UvrB is unable to bend DNA.

Duke Scholars

Published In

J Mol Biol

DOI

ISSN

0022-2836

Publication Date

September 2, 1994

Volume

241

Issue

5

Start / End Page

645 / 650

Location

Netherlands

Related Subject Headings

  • Ultraviolet Rays
  • Spectrophotometry, Ultraviolet
  • Nucleic Acid Conformation
  • Mutation
  • Microscopy, Electron
  • Escherichia coli Proteins
  • Endodeoxyribonucleases
  • DNA-Binding Proteins
  • DNA Helicases
  • DNA
 

Citation

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Hsu, D. S., Takahashi, M., Delagoutte, E., Bertrand-Burggraf, E., Wang, Y. H., Norden, B., … Sancar, A. (1994). Flow linear dichroism and electron microscopic analysis of protein-DNA complexes of a mutant UvrB protein that binds to but cannot kink DNA. J Mol Biol, 241(5), 645–650. https://doi.org/10.1006/jmbi.1994.1541
Hsu, D. S., M. Takahashi, E. Delagoutte, E. Bertrand-Burggraf, Y. H. Wang, B. Norden, R. P. Fuchs, J. Griffith, and A. Sancar. “Flow linear dichroism and electron microscopic analysis of protein-DNA complexes of a mutant UvrB protein that binds to but cannot kink DNA.J Mol Biol 241, no. 5 (September 2, 1994): 645–50. https://doi.org/10.1006/jmbi.1994.1541.
Hsu DS, Takahashi M, Delagoutte E, Bertrand-Burggraf E, Wang YH, Norden B, et al. Flow linear dichroism and electron microscopic analysis of protein-DNA complexes of a mutant UvrB protein that binds to but cannot kink DNA. J Mol Biol. 1994 Sep 2;241(5):645–50.
Hsu, D. S., et al. “Flow linear dichroism and electron microscopic analysis of protein-DNA complexes of a mutant UvrB protein that binds to but cannot kink DNA.J Mol Biol, vol. 241, no. 5, Sept. 1994, pp. 645–50. Pubmed, doi:10.1006/jmbi.1994.1541.
Hsu DS, Takahashi M, Delagoutte E, Bertrand-Burggraf E, Wang YH, Norden B, Fuchs RP, Griffith J, Sancar A. Flow linear dichroism and electron microscopic analysis of protein-DNA complexes of a mutant UvrB protein that binds to but cannot kink DNA. J Mol Biol. 1994 Sep 2;241(5):645–650.
Journal cover image

Published In

J Mol Biol

DOI

ISSN

0022-2836

Publication Date

September 2, 1994

Volume

241

Issue

5

Start / End Page

645 / 650

Location

Netherlands

Related Subject Headings

  • Ultraviolet Rays
  • Spectrophotometry, Ultraviolet
  • Nucleic Acid Conformation
  • Mutation
  • Microscopy, Electron
  • Escherichia coli Proteins
  • Endodeoxyribonucleases
  • DNA-Binding Proteins
  • DNA Helicases
  • DNA