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- Title
NMR Characterization of the DNA-binding Domain of Arabidopsis thaliana Telomere Repeat Factor.
- Authors
Lee, Inhwan; Kim, Heeyoun; Ko, Yoon-Joo; Lee, Weontae
- Abstract
Telomere repeat factor ( AtTRF) derived from Arabidopsis thaliana contains a myb-like domain that binds to double-stranded telomeric DNA. We cloned the myb-like domain of AtTRF ( AtTRFmyb ) into a pET-15b vector and expressed the protein in Escherichia coli. AtTRFmyb was purified using Ni-affinity chromatography. DNA-binding mode has been examined by electrophoretic mobility shift assay ( EMSA). Fluorescence-quenching experiments determined a Kd value of 6.62 nM between AtTRFmyb and plant telomeric DNA. Data from nuclear magnetic resonance ( NMR) spectroscopy together with TALOS+ program provide the secondary structures of AtTRFmyb , suggesting that AtTRFmyb binds its plant telomere DNA with three α-helices of a DNA-binding motif.
- Subjects
NUCLEAR magnetic resonance; DNA-binding proteins; ARABIDOPSIS; CHROMATOGRAPHIC analysis; ELECTROPHORETIC deposition
- Publication
Bulletin of the Korean Chemical Society, 2016, Vol 37, Issue 4, p485
- ISSN
0253-2964
- Publication type
Article
- DOI
10.1002/bkcs.10708