Sequence-specific and DNA structure-dependent interactions of Escherichia coli MutS and human p53 with DNA

Cobb, Andrew M., Jackson, Brian R., Kim, Ella, Bond, Philip L. and Bowater, Richard P. (2013) Sequence-specific and DNA structure-dependent interactions of Escherichia coli MutS and human p53 with DNA. Analytical Biochemistry, 442 1: 51-61. doi:10.1016/j.ab.2013.07.033


Author Cobb, Andrew M.
Jackson, Brian R.
Kim, Ella
Bond, Philip L.
Bowater, Richard P.
Title Sequence-specific and DNA structure-dependent interactions of Escherichia coli MutS and human p53 with DNA
Formatted title
Sequence-specific and DNA structure-dependent interactions of Escherichia coli MutS and human p53 with DNA
Journal name Analytical Biochemistry   Check publisher's open access policy
ISSN 0003-2697
1096-0309
Publication date 2013-11-01
Sub-type Article (original research)
DOI 10.1016/j.ab.2013.07.033
Open Access Status Not Open Access
Volume 442
Issue 1
Start page 51
End page 61
Total pages 11
Place of publication Philadelphia, PA, United States
Publisher Elsevier
Language eng
Formatted abstract
Many proteins involved in DNA repair systems interact with DNA that has structure altered from the typical B-form helix. Using magnetic beads to immobilize DNAs containing various types of structures, we evaluated the in vitro binding activities of two well-characterized DNA repair proteins, Escherichia coli MutS and human p53. E. coli MutS bound to double-stranded DNAs, with higher affinity for a G/T mismatch compared to a G/A mismatch and highest affinity for larger non-B-DNA structures. E. coli MutS bound best to DNA between pH 6 and 9. Experiments discriminated between modes of p53-DNA binding, and increasing ionic strength reduced p53 binding to nonspecific double-stranded DNA, but had minor effects on binding to consensus response sequences or single-stranded DNA. Compared to nonspecific DNA sequences, p53 bound with a higher affinity to mismatches and base insertions, while binding to various hairpin structures was similar to that observed to its consensus DNA sequence. For hairpins containing CTG repeats, the extent of p53 binding was proportional to the size of the repeat. In summary, using the flexibility of the magnetic bead separation assay we demonstrate that pH and ionic strength influence the binding of two DNA repair proteins to a variety of DNA structures.
Keyword Biotin-streptavidin
Immobilized DNA substrates
MutS
P53
Protein-DNA binding
Unusual DNA structures
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2014 Collection
Advanced Water Management Centre Publications
 
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