Journal article
DNA repair in a protein-DNA complex: searching for the key to get in
Mutation research, Vol.577(1-2), pp.118-130
09/04/2005
Handle:
https://hdl.handle.net/2376/116027
PMID: 15913668
Abstract
An obstacle encountered by nucleotide excision repair (NER) proteins during repair of the genome is the masking of bulky lesions by DNA binding proteins. For example, certain transcription factors are known to be impediments, and suppress damage removal at their recognition sequences. We have used well-defined protein-DNA complexes to study the molecular mechanism(s) used by repair proteins in gaining access to DNA lesions in chromatin. Using transcription factor IIIA (TFIIIA) and the 5S ribosomal RNA gene (5S rDNA), we previously measured position-dependent effects of cyclobutane pyrimidine dimers (CPDs) at five different sites within the internal control region (ICR) on complex formation [Y. Kwon, M.J. Smerdon, Binding of zinc finger protein transcription factor IIIA to its cognate DNA sequence with single UV photoproducts at specific sites and its effect on DNA repair, J. Biol. Chem. 278 (2003) 45451-45459]. We found that CPDs at two of these sites enhance the TFIIIA-rDNA dissociation rate, which correlates with enhanced repair at these two sites. Here, we used a novel approach to directly compare dissociation of randomly damaged rDNA with NER. We refined the relationship between dissociation and repair of the complex by examining all CPD sites in the transcribed strand. A 214 bp 5S rDNA fragment was irradiated with UV light to produce CPDs at dipyrimidine sites and approximately 1 CPD per fragment. Positions of CPDs that alter binding of TFIIIA were determined by T4 endonuclease V mapping of TFIIIA-bound and unbound fractions of UV-irradiated DNA. As expected, the results reveal that dissociation of TFIIIA from the complex is significantly enhanced by CPDs within the ICR. Moreover, the levels of dissociation induced by CPDs were quantitatively compared with their repair efficiency, and indicate that repair rates of most CPDs in the complex closely correlate with the dissociation rates. In addition, changes in dissociation rate are similar to changes in CPD frequency induced by TFIIIA binding. These findings indicate that structural compatibility of a DNA lesion within a protein-DNA complex can determine both lesion frequency and repair efficiency.
Metrics
14 Record Views
Details
- Title
- DNA repair in a protein-DNA complex: searching for the key to get in
- Creators
- Youngho Kwon - Biochemistry and Biophysics, School of Molecular Biosciences, Washington State University, Pullman, WA 99164-4660, USAMichael J Smerdon
- Publication Details
- Mutation research, Vol.577(1-2), pp.118-130
- Academic Unit
- Molecular Biosciences, School of
- Publisher
- Netherlands
- Grant note
- R37 ES002614-25 / NIEHS NIH HHS ES02614 / NIEHS NIH HHS
- Identifiers
- 99900548177701842
- Language
- English
- Resource Type
- Journal article