Cisplatin-DNA binding specificity of calf high-mobility group 1 protein

John Turchi, Ming Li, Karen M. Henkels

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

We have identified a series of proteins with an affinity for cisplatin- damaged DNA using damaged DNA affinity chromatography. We have purified one of these proteins to homogeneity on the basis of a mobility shift assay detecting binding to cisplatin-damaged DNA. The protein was identified as high-mobility group 1 protein (HMG-1) by N-terminal protein sequence analysis. Analysis of a variety of DNA structures revealed that fully duplex DNAs were the best substrates for HMG-1 binding, while partial duplexes were less avidly bound. The decreased levels of binding are attributed to the length of the duplex region of the DNA substrates. A 3-fold increase in binding was observed when a cisplatin-damaged DNA substrate containing a single break in the phosphodiester backbone was joined by DNA ligase. The strict DNA size dependence of binding was also assessed, and a 10-fold increase in binding was observed when the length of the DNA duplex was increased from 44 to 180 base pairs (bp) at the same level of cisplatin damage. HMG-1 binding also was correlated with the degree of cisplatin-DNA damage, suggesting a higher affinity for DNA containing multiple cisplatin adducts. Nuclease degradation of the cisplatin-damaged DNA demonstrated that at the lowest levels of cisplatin damage all of the substrates contained at least one cisplatin adduct. The potential role of HMG-1 in the repair of cisplatin-DNA adducts is discussed.

Original languageEnglish (US)
Pages (from-to)2992-3000
Number of pages9
JournalBiochemistry
Volume35
Issue number9
DOIs
StatePublished - Mar 5 1996
Externally publishedYes

Fingerprint

High Mobility Group Proteins
Cisplatin
DNA
Proteins
Protein Binding
Substrates
DNA Ligases
Affinity chromatography
Protein Sequence Analysis
Electrophoretic Mobility Shift Assay
Affinity Chromatography
Base Pairing
DNA Damage
Assays

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cisplatin-DNA binding specificity of calf high-mobility group 1 protein. / Turchi, John; Li, Ming; Henkels, Karen M.

In: Biochemistry, Vol. 35, No. 9, 05.03.1996, p. 2992-3000.

Research output: Contribution to journalArticle

Turchi, John ; Li, Ming ; Henkels, Karen M. / Cisplatin-DNA binding specificity of calf high-mobility group 1 protein. In: Biochemistry. 1996 ; Vol. 35, No. 9. pp. 2992-3000.
@article{922073fc68214c4faf42ffdf3f50d1b7,
title = "Cisplatin-DNA binding specificity of calf high-mobility group 1 protein",
abstract = "We have identified a series of proteins with an affinity for cisplatin- damaged DNA using damaged DNA affinity chromatography. We have purified one of these proteins to homogeneity on the basis of a mobility shift assay detecting binding to cisplatin-damaged DNA. The protein was identified as high-mobility group 1 protein (HMG-1) by N-terminal protein sequence analysis. Analysis of a variety of DNA structures revealed that fully duplex DNAs were the best substrates for HMG-1 binding, while partial duplexes were less avidly bound. The decreased levels of binding are attributed to the length of the duplex region of the DNA substrates. A 3-fold increase in binding was observed when a cisplatin-damaged DNA substrate containing a single break in the phosphodiester backbone was joined by DNA ligase. The strict DNA size dependence of binding was also assessed, and a 10-fold increase in binding was observed when the length of the DNA duplex was increased from 44 to 180 base pairs (bp) at the same level of cisplatin damage. HMG-1 binding also was correlated with the degree of cisplatin-DNA damage, suggesting a higher affinity for DNA containing multiple cisplatin adducts. Nuclease degradation of the cisplatin-damaged DNA demonstrated that at the lowest levels of cisplatin damage all of the substrates contained at least one cisplatin adduct. The potential role of HMG-1 in the repair of cisplatin-DNA adducts is discussed.",
author = "John Turchi and Ming Li and Henkels, {Karen M.}",
year = "1996",
month = "3",
day = "5",
doi = "10.1021/bi951843j",
language = "English (US)",
volume = "35",
pages = "2992--3000",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "9",

}

TY - JOUR

T1 - Cisplatin-DNA binding specificity of calf high-mobility group 1 protein

AU - Turchi, John

AU - Li, Ming

AU - Henkels, Karen M.

PY - 1996/3/5

Y1 - 1996/3/5

N2 - We have identified a series of proteins with an affinity for cisplatin- damaged DNA using damaged DNA affinity chromatography. We have purified one of these proteins to homogeneity on the basis of a mobility shift assay detecting binding to cisplatin-damaged DNA. The protein was identified as high-mobility group 1 protein (HMG-1) by N-terminal protein sequence analysis. Analysis of a variety of DNA structures revealed that fully duplex DNAs were the best substrates for HMG-1 binding, while partial duplexes were less avidly bound. The decreased levels of binding are attributed to the length of the duplex region of the DNA substrates. A 3-fold increase in binding was observed when a cisplatin-damaged DNA substrate containing a single break in the phosphodiester backbone was joined by DNA ligase. The strict DNA size dependence of binding was also assessed, and a 10-fold increase in binding was observed when the length of the DNA duplex was increased from 44 to 180 base pairs (bp) at the same level of cisplatin damage. HMG-1 binding also was correlated with the degree of cisplatin-DNA damage, suggesting a higher affinity for DNA containing multiple cisplatin adducts. Nuclease degradation of the cisplatin-damaged DNA demonstrated that at the lowest levels of cisplatin damage all of the substrates contained at least one cisplatin adduct. The potential role of HMG-1 in the repair of cisplatin-DNA adducts is discussed.

AB - We have identified a series of proteins with an affinity for cisplatin- damaged DNA using damaged DNA affinity chromatography. We have purified one of these proteins to homogeneity on the basis of a mobility shift assay detecting binding to cisplatin-damaged DNA. The protein was identified as high-mobility group 1 protein (HMG-1) by N-terminal protein sequence analysis. Analysis of a variety of DNA structures revealed that fully duplex DNAs were the best substrates for HMG-1 binding, while partial duplexes were less avidly bound. The decreased levels of binding are attributed to the length of the duplex region of the DNA substrates. A 3-fold increase in binding was observed when a cisplatin-damaged DNA substrate containing a single break in the phosphodiester backbone was joined by DNA ligase. The strict DNA size dependence of binding was also assessed, and a 10-fold increase in binding was observed when the length of the DNA duplex was increased from 44 to 180 base pairs (bp) at the same level of cisplatin damage. HMG-1 binding also was correlated with the degree of cisplatin-DNA damage, suggesting a higher affinity for DNA containing multiple cisplatin adducts. Nuclease degradation of the cisplatin-damaged DNA demonstrated that at the lowest levels of cisplatin damage all of the substrates contained at least one cisplatin adduct. The potential role of HMG-1 in the repair of cisplatin-DNA adducts is discussed.

UR - http://www.scopus.com/inward/record.url?scp=0029868834&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029868834&partnerID=8YFLogxK

U2 - 10.1021/bi951843j

DO - 10.1021/bi951843j

M3 - Article

C2 - 8608137

AN - SCOPUS:0029868834

VL - 35

SP - 2992

EP - 3000

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 9

ER -