NMR characterization of the DNA-binding domain of arabidopsis thaliana telomere repeat factor

Inhwan Lee, Heeyoun Kim, Yoon Joo Ko, Weontae Lee

Research output: Contribution to journalArticle

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 chro-matography. 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 tel-omeric 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.

Original languageEnglish
Pages (from-to)485-489
Number of pages5
JournalBulletin of the Korean Chemical Society
Volume37
Issue number4
DOIs
Publication statusPublished - 2016 Apr 1

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

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