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

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

doi:10.1002/bkcs.10708

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 journal › Article › peer-review

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 (AtTRF^myb) into a pET-15b vector and expressed the protein in Escherichia coli. AtTRF^myb was purified using Ni-affinity chro-matography. 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 tel-omeric 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.

Original language	English
Pages (from-to)	485-489
Number of pages	5
Journal	Bulletin of the Korean Chemical Society
Volume	37
Issue number	4
DOIs	https://doi.org/10.1002/bkcs.10708
Publication status	Published - 2016 Apr 1

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© 2016 Korean Chemical Society, Seoul & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

All Science Journal Classification (ASJC) codes

Chemistry(all)

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10.1002/bkcs.10708

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title = "NMR characterization of the DNA-binding domain of arabidopsis thaliana telomere repeat factor",

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.",

author = "Inhwan Lee and Heeyoun Kim and Ko, {Yoon Joo} and Weontae Lee",

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T1 - NMR characterization of the DNA-binding domain of arabidopsis thaliana telomere repeat factor

AU - Lee, Inhwan

AU - Kim, Heeyoun

AU - Ko, Yoon Joo

AU - Lee, Weontae

PY - 2016/4/1

Y1 - 2016/4/1

N2 - 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.

AB - 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.

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

Abstract

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