Reevaluation of a dna binding activity with specificity for chemically modified dna

Robin H. Chou; Moon Shong Tang; Steven M. Cohn; Michael W. Lieberman

doi:10.1093/carcin/3.6.719

Reevaluation of a dna binding activity with specificity for chemically modified dna

Robin H. Chou, Moon Shong Tang, Steven M. Cohn, Michael W. Lieberman

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

1 Scopus citations

Abstract

A DNA binding activity which appeared in direct filter binding assays to show specificity for DNA modified by N-acetoxy-2-acetylaminofluorene (AAF), N-methyl-N-nitro-sourea and methylmethanesulfonate (Moranelli and Lieber-man, Proc. Natl. Acad. Sci. USA, 77, 3201, 1980) has been further purified and characterized. Equilibrium competition binding experiments indicate this activity shows little, or no, preferential binding of AAF-modified DNA compared to single-stranded DNA and only a small preference compared to u.v.-irradiated DNA or double-stranded DNA. These findings demonstrate the difficulty of using direct filter binding assays when purifying proteins with a desired biological function and the need to perform competition assays when evaluating specificity.

Original language	English (US)
Pages (from-to)	719-721
Number of pages	3
Journal	Carcinogenesis
Volume	3
Issue number	6
DOIs	https://doi.org/10.1093/carcin/3.6.719
State	Published - 1982
Externally published	Yes

ASJC Scopus subject areas

Cancer Research

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10.1093/carcin/3.6.719

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@article{3b6f8b5cf40b4ebe9d73a929d01dbb68,

title = "Reevaluation of a dna binding activity with specificity for chemically modified dna",

abstract = "A DNA binding activity which appeared in direct filter binding assays to show specificity for DNA modified by N-acetoxy-2-acetylaminofluorene (AAF), N-methyl-N-nitro-sourea and methylmethanesulfonate (Moranelli and Lieber-man, Proc. Natl. Acad. Sci. USA, 77, 3201, 1980) has been further purified and characterized. Equilibrium competition binding experiments indicate this activity shows little, or no, preferential binding of AAF-modified DNA compared to single-stranded DNA and only a small preference compared to u.v.-irradiated DNA or double-stranded DNA. These findings demonstrate the difficulty of using direct filter binding assays when purifying proteins with a desired biological function and the need to perform competition assays when evaluating specificity.",

author = "Chou, {Robin H.} and Tang, {Moon Shong} and Cohn, {Steven M.} and Lieberman, {Michael W.}",

note = "Funding Information: We thank Steven L. Dresler, Dard J. Hunting and Michael B. Kastan for their thoughtful comments. This work was supported by NIH Grant CA 20513 and the following companies: Brown & Williamson Tobacco Corporation; Philip Morris Incorporated; R.J. Reynolds Tobacco Company; and United States Tobacco Company. RHC and SMC were supported by NIH Training Grant ES 07066, and MST was supported by NIH Training Grant CAO9118.",

year = "1982",

doi = "10.1093/carcin/3.6.719",

language = "English (US)",

volume = "3",

pages = "719--721",

journal = "Carcinogenesis",

issn = "0143-3334",

publisher = "Oxford University Press",

number = "6",

}

TY - JOUR

T1 - Reevaluation of a dna binding activity with specificity for chemically modified dna

AU - Chou, Robin H.

AU - Tang, Moon Shong

AU - Cohn, Steven M.

AU - Lieberman, Michael W.

N1 - Funding Information: We thank Steven L. Dresler, Dard J. Hunting and Michael B. Kastan for their thoughtful comments. This work was supported by NIH Grant CA 20513 and the following companies: Brown & Williamson Tobacco Corporation; Philip Morris Incorporated; R.J. Reynolds Tobacco Company; and United States Tobacco Company. RHC and SMC were supported by NIH Training Grant ES 07066, and MST was supported by NIH Training Grant CAO9118.

PY - 1982

Y1 - 1982

N2 - A DNA binding activity which appeared in direct filter binding assays to show specificity for DNA modified by N-acetoxy-2-acetylaminofluorene (AAF), N-methyl-N-nitro-sourea and methylmethanesulfonate (Moranelli and Lieber-man, Proc. Natl. Acad. Sci. USA, 77, 3201, 1980) has been further purified and characterized. Equilibrium competition binding experiments indicate this activity shows little, or no, preferential binding of AAF-modified DNA compared to single-stranded DNA and only a small preference compared to u.v.-irradiated DNA or double-stranded DNA. These findings demonstrate the difficulty of using direct filter binding assays when purifying proteins with a desired biological function and the need to perform competition assays when evaluating specificity.

AB - A DNA binding activity which appeared in direct filter binding assays to show specificity for DNA modified by N-acetoxy-2-acetylaminofluorene (AAF), N-methyl-N-nitro-sourea and methylmethanesulfonate (Moranelli and Lieber-man, Proc. Natl. Acad. Sci. USA, 77, 3201, 1980) has been further purified and characterized. Equilibrium competition binding experiments indicate this activity shows little, or no, preferential binding of AAF-modified DNA compared to single-stranded DNA and only a small preference compared to u.v.-irradiated DNA or double-stranded DNA. These findings demonstrate the difficulty of using direct filter binding assays when purifying proteins with a desired biological function and the need to perform competition assays when evaluating specificity.

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DO - 10.1093/carcin/3.6.719

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VL - 3

SP - 719

EP - 721

JO - Carcinogenesis

JF - Carcinogenesis

IS - 6

ER -

Reevaluation of a dna binding activity with specificity for chemically modified dna

Abstract

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