Journal article
Single-molecule analysis reveals differential effect of ssDNA-binding proteins on DNA translocation by XPD helicase
Molecular cell, Vol.35(5), pp.694-703
09/11/2009
DOI: 10.1016/j.molcel.2009.07.003
PMCID: PMC2776038
PMID: 19748362
Abstract
An encounter between a DNA translocating enzyme and a DNA-bound protein must occur frequently in the cell but little is known about its outcome. Here, we developed a multi-color single molecule fluorescence approach to simultaneously monitor single stranded (ss) DNA translocation by a helicase and the fate of another protein bound to the same DNA. Distance-dependent fluorescence quenching by the iron-sulfur cluster of the archaeal XPD (Rad3) helicase was used as a calibrated proximity signal. Despite the similar equilibrium DNA binding properties, the two cognate ssDNA binding proteins, RPA1 and RPA2, differentially affected XPD translocation. RPA1 competed with XPD for ssDNA access. In contrast, RPA2 did not interfere with XPD-ssDNA binding but markedly slowed down XPD translocation. Mechanistic models of bypassing DNA-bound proteins by the Rad3 family helicases and their biological implications are discussed.
Details
- Title: Subtitle
- Single-molecule analysis reveals differential effect of ssDNA-binding proteins on DNA translocation by XPD helicase
- Creators
- Masayoshi Honda - Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, 61801 USAJeehae Park - Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois, 61801 USARobert A Pugh - Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, 61801 USATaekjip Ha - Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois, 61801 USAMaria Spies - Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, 61801 USA
- Resource Type
- Journal article
- Publication Details
- Molecular cell, Vol.35(5), pp.694-703
- DOI
- 10.1016/j.molcel.2009.07.003
- PMID
- 19748362
- PMCID
- PMC2776038
- ISSN
- 1097-2765
- eISSN
- 1097-4164
- Grant note
- || HHMI_ / Howard Hughes Medical Institute :
- Language
- English
- Date published
- 09/11/2009
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984025283302771
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