NMR  Characterization of the  DNA ‐binding Domain of  Arabidopsis thaliana  Telomere Repeat Factor | Zendy

Lee Inhwan | Zendy; Kim Heeyoun | Zendy; Ko YoonJoo | Zendy; Lee Weontae | Zendy

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NMR Characterization of the DNA ‐binding Domain of Arabidopsis thaliana Telomere Repeat Factor

Author(s) -

Lee Inhwan,

Kim Heeyoun,

Ko YoonJoo,

Lee Weontae

Publication year - 2016

Publication title -

bulletin of the korean chemical society

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.237

H-Index - 59

ISSN - 1229-5949

DOI - 10.1002/bkcs.10708

Subject(s) - telomere , myb , dna binding domain , telomere binding protein , dna , electrophoretic mobility shift assay , arabidopsis thaliana , microbiology and biotechnology , dna binding protein , chemistry , biology , transcription factor , biochemistry , gene , mutant

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 ( AtTRF myb ) into a pET ‐15b vector and expressed the protein in Escherichia coli . AtTRF myb was purified using Ni‐affinity chromatography. DNA ‐binding mode has been examined by electrophoretic mobility shift assay ( EMSA ). Fluorescence‐quenching experiments determined a K d value of 6.62 nM between AtTRF myb and plant telomeric DNA . Data from nuclear magnetic resonance ( NMR ) spectroscopy together with TALOS + program provide the secondary structures of AtTRF myb , suggesting that AtTRF myb binds its plant telomere DNA with three α‐helices of a DNA ‐binding motif.

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