NMR and fluorescence studies of DNA binding domain of INI1/hSNF5

Dongju Lee; Sunjin Moon; Jihye Yun; Eunhee Kim; Chaejoon Cheong; Weontae Lee

doi:10.5012/bkcs.2014.35.9.2753

NMR and fluorescence studies of DNA binding domain of INI1/hSNF5

Dongju Lee, Sunjin Moon, Jihye Yun, Eunhee Kim, Chaejoon Cheong, Weontae Lee

Research output: Contribution to journal › Article › peer-review

Abstract

INtegrase Interactor 1 protein (INI1/hSNF5) or BRG1-associated factor 47 (BAF47) is a SWI/SNF-related matrix associated actin dependent regulator of chromatin subfamily B member. DNA binding domain of INI1/hSNF5 is cloned into E.coli expression vectors, pET32a and purified as a monomer using size exclusion chromatography. NMR data show that INI1^DBD has folded state with high population of α-helices. By fluorescence-quenching experiments, binding affinities between INI1^DBD and two double stranded DNA fragments were determined as 29.9 ± 2.6 μM (GAL4-1) and 258.7 ± 5.8 (GAL4-2) μM, respectively. Our data revealed that DNA binding domain of INI1/hSNF5 binds to transcriptional DNA sequences, and it could play an important role as a transcriptional regulator.

Original language	English
Pages (from-to)	2753-2757
Number of pages	5
Journal	Bulletin of the Korean Chemical Society
Volume	35
Issue number	9
DOIs	https://doi.org/10.5012/bkcs.2014.35.9.2753
Publication status	Published - 2014 Sept 20

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© 2014, Korean Chemical Society. All rights reserved.

All Science Journal Classification (ASJC) codes

Chemistry(all)

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10.5012/bkcs.2014.35.9.2753

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title = "NMR and fluorescence studies of DNA binding domain of INI1/hSNF5",

abstract = "INtegrase Interactor 1 protein (INI1/hSNF5) or BRG1-associated factor 47 (BAF47) is a SWI/SNF-related matrix associated actin dependent regulator of chromatin subfamily B member. DNA binding domain of INI1/hSNF5 is cloned into E.coli expression vectors, pET32a and purified as a monomer using size exclusion chromatography. NMR data show that INI1DBD has folded state with high population of α-helices. By fluorescence-quenching experiments, binding affinities between INI1DBD and two double stranded DNA fragments were determined as 29.9 ± 2.6 μM (GAL4-1) and 258.7 ± 5.8 (GAL4-2) μM, respectively. Our data revealed that DNA binding domain of INI1/hSNF5 binds to transcriptional DNA sequences, and it could play an important role as a transcriptional regulator.",

author = "Dongju Lee and Sunjin Moon and Jihye Yun and Eunhee Kim and Chaejoon Cheong and Weontae Lee",

year = "2014",

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T1 - NMR and fluorescence studies of DNA binding domain of INI1/hSNF5

AU - Lee, Dongju

AU - Moon, Sunjin

AU - Yun, Jihye

AU - Kim, Eunhee

AU - Cheong, Chaejoon

AU - Lee, Weontae

PY - 2014/9/20

Y1 - 2014/9/20

N2 - INtegrase Interactor 1 protein (INI1/hSNF5) or BRG1-associated factor 47 (BAF47) is a SWI/SNF-related matrix associated actin dependent regulator of chromatin subfamily B member. DNA binding domain of INI1/hSNF5 is cloned into E.coli expression vectors, pET32a and purified as a monomer using size exclusion chromatography. NMR data show that INI1DBD has folded state with high population of α-helices. By fluorescence-quenching experiments, binding affinities between INI1DBD and two double stranded DNA fragments were determined as 29.9 ± 2.6 μM (GAL4-1) and 258.7 ± 5.8 (GAL4-2) μM, respectively. Our data revealed that DNA binding domain of INI1/hSNF5 binds to transcriptional DNA sequences, and it could play an important role as a transcriptional regulator.

AB - INtegrase Interactor 1 protein (INI1/hSNF5) or BRG1-associated factor 47 (BAF47) is a SWI/SNF-related matrix associated actin dependent regulator of chromatin subfamily B member. DNA binding domain of INI1/hSNF5 is cloned into E.coli expression vectors, pET32a and purified as a monomer using size exclusion chromatography. NMR data show that INI1DBD has folded state with high population of α-helices. By fluorescence-quenching experiments, binding affinities between INI1DBD and two double stranded DNA fragments were determined as 29.9 ± 2.6 μM (GAL4-1) and 258.7 ± 5.8 (GAL4-2) μM, respectively. Our data revealed that DNA binding domain of INI1/hSNF5 binds to transcriptional DNA sequences, and it could play an important role as a transcriptional regulator.

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JO - Bulletin of the Korean Chemical Society

JF - Bulletin of the Korean Chemical Society

IS - 9

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NMR and fluorescence studies of DNA binding domain of INI1/hSNF5

Abstract

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