Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases

Moon Shong Tang, Chong Soon Lee, Richard Doisy, Lisa Ross, Donald R. Needham-VanDevanter, Laurence Hurley

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

The recognition and repair of the helix-stabilizing and relatively nondistortive CC-1065-(N3-adenine)-DNA adduct by UVRABC nuclease has been investigated both in vivo with ΦX174 RFI DNA by a transfection assay and in vitro by a site-directed adduct in a 117 base pair fragment from M13mpl. CC-1065 is a potent antitumor antibiotic produced by Streptomyces zelensis which binds within the minor groove of DNA through N3 of adenine. In contrast to the helix-destabilizing and distortive modifications of DNA caused by ultraviolet light or N-acetoxy-2-(acetylamino)fluorene, CC-1065 increases the melting point of DNA and decreases the S1 nuclease activity. Using a viral DNA-Escherichia coli transfection system, we have found that the uvrA, uvrB, and uvrC genes, which code for the major excision repair proteins for UV- and NAAAF-induced DNA damage, are also involved in the repair of CC-1065-DNA adducts. In contrast, the uvrD gene product, which has been found to be involved in the repair of UV damage, has no effect in repairing CC-1065-DNA adducts. Purified UVRA, UVRB, and UVRC proteins must work in concert to incise the drug-modified ΦX174 RFI DNA. Using a site-directed and multiple CC-1065 modified (MspI-BstNI) 117 base pair fragment from M13mpl, we have found that UVRABC nuclease incises at the eighth phosphodiester bond on the 5′ side of the CC-1065-DNA adduct on the drug-modified strand. The enzymes do not cut the noncovalently modified strand. At low drug binding ratios, of the four CC-1065 binding sites identified in the (MspI-BstNI) 117 base pair fragment, GATTA*, GGAAA*, GATAA*, and TTTTA* (* indicates the covalently modified adenine), only the adduct at the high-affinity binding site, GATTA*, is incised by the UVRABC nucleases. No difference in the effect of CC-1065 on local DNA structure, as determined by the DNase I cleavage pattern, was evident among these sites. At high drug binding ratios, a fifth drug binding site, AGCTA*, is identified. At this concentration UVRABC nucleases are unable to incise any of these five CC-1065-DNA adducts. The DNA sequence and/or helix-stabilizing effect of multiple adducts may determine the recognition and/or incision of the drug-DNA adduct by UVRABC nuclease. These results are discussed in relation to the structure of the CC-1065-DNA adduct and the effect of drug binding on local DNA structure.

Original languageEnglish (US)
Pages (from-to)893-901
Number of pages9
JournalBiochemistry
Volume27
Issue number3
StatePublished - 1988
Externally publishedYes

Fingerprint

CC 1065
DNA Adducts
Adenine
Repair
DNA
Pharmaceutical Preparations
Base Pairing
Binding Sites
Transfection
Nucleic Acid Denaturation
Genes
Deoxyribonuclease I
Viral DNA
Streptomyces
Ultraviolet Rays
DNA sequences
DNA Repair
DNA Damage
Escherichia coli
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Tang, M. S., Lee, C. S., Doisy, R., Ross, L., Needham-VanDevanter, D. R., & Hurley, L. (1988). Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases. Biochemistry, 27(3), 893-901.

Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases. / Tang, Moon Shong; Lee, Chong Soon; Doisy, Richard; Ross, Lisa; Needham-VanDevanter, Donald R.; Hurley, Laurence.

In: Biochemistry, Vol. 27, No. 3, 1988, p. 893-901.

Research output: Contribution to journalArticle

Tang, MS, Lee, CS, Doisy, R, Ross, L, Needham-VanDevanter, DR & Hurley, L 1988, 'Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases', Biochemistry, vol. 27, no. 3, pp. 893-901.
Tang MS, Lee CS, Doisy R, Ross L, Needham-VanDevanter DR, Hurley L. Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases. Biochemistry. 1988;27(3):893-901.
Tang, Moon Shong ; Lee, Chong Soon ; Doisy, Richard ; Ross, Lisa ; Needham-VanDevanter, Donald R. ; Hurley, Laurence. / Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases. In: Biochemistry. 1988 ; Vol. 27, No. 3. pp. 893-901.
@article{e8d87101250f47ebbc48defafb56cfdd,
title = "Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases",
abstract = "The recognition and repair of the helix-stabilizing and relatively nondistortive CC-1065-(N3-adenine)-DNA adduct by UVRABC nuclease has been investigated both in vivo with ΦX174 RFI DNA by a transfection assay and in vitro by a site-directed adduct in a 117 base pair fragment from M13mpl. CC-1065 is a potent antitumor antibiotic produced by Streptomyces zelensis which binds within the minor groove of DNA through N3 of adenine. In contrast to the helix-destabilizing and distortive modifications of DNA caused by ultraviolet light or N-acetoxy-2-(acetylamino)fluorene, CC-1065 increases the melting point of DNA and decreases the S1 nuclease activity. Using a viral DNA-Escherichia coli transfection system, we have found that the uvrA, uvrB, and uvrC genes, which code for the major excision repair proteins for UV- and NAAAF-induced DNA damage, are also involved in the repair of CC-1065-DNA adducts. In contrast, the uvrD gene product, which has been found to be involved in the repair of UV damage, has no effect in repairing CC-1065-DNA adducts. Purified UVRA, UVRB, and UVRC proteins must work in concert to incise the drug-modified ΦX174 RFI DNA. Using a site-directed and multiple CC-1065 modified (MspI-BstNI) 117 base pair fragment from M13mpl, we have found that UVRABC nuclease incises at the eighth phosphodiester bond on the 5′ side of the CC-1065-DNA adduct on the drug-modified strand. The enzymes do not cut the noncovalently modified strand. At low drug binding ratios, of the four CC-1065 binding sites identified in the (MspI-BstNI) 117 base pair fragment, GATTA*, GGAAA*, GATAA*, and TTTTA* (* indicates the covalently modified adenine), only the adduct at the high-affinity binding site, GATTA*, is incised by the UVRABC nucleases. No difference in the effect of CC-1065 on local DNA structure, as determined by the DNase I cleavage pattern, was evident among these sites. At high drug binding ratios, a fifth drug binding site, AGCTA*, is identified. At this concentration UVRABC nucleases are unable to incise any of these five CC-1065-DNA adducts. The DNA sequence and/or helix-stabilizing effect of multiple adducts may determine the recognition and/or incision of the drug-DNA adduct by UVRABC nuclease. These results are discussed in relation to the structure of the CC-1065-DNA adduct and the effect of drug binding on local DNA structure.",
author = "Tang, {Moon Shong} and Lee, {Chong Soon} and Richard Doisy and Lisa Ross and Needham-VanDevanter, {Donald R.} and Laurence Hurley",
year = "1988",
language = "English (US)",
volume = "27",
pages = "893--901",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "3",

}

TY - JOUR

T1 - Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases

AU - Tang, Moon Shong

AU - Lee, Chong Soon

AU - Doisy, Richard

AU - Ross, Lisa

AU - Needham-VanDevanter, Donald R.

AU - Hurley, Laurence

PY - 1988

Y1 - 1988

N2 - The recognition and repair of the helix-stabilizing and relatively nondistortive CC-1065-(N3-adenine)-DNA adduct by UVRABC nuclease has been investigated both in vivo with ΦX174 RFI DNA by a transfection assay and in vitro by a site-directed adduct in a 117 base pair fragment from M13mpl. CC-1065 is a potent antitumor antibiotic produced by Streptomyces zelensis which binds within the minor groove of DNA through N3 of adenine. In contrast to the helix-destabilizing and distortive modifications of DNA caused by ultraviolet light or N-acetoxy-2-(acetylamino)fluorene, CC-1065 increases the melting point of DNA and decreases the S1 nuclease activity. Using a viral DNA-Escherichia coli transfection system, we have found that the uvrA, uvrB, and uvrC genes, which code for the major excision repair proteins for UV- and NAAAF-induced DNA damage, are also involved in the repair of CC-1065-DNA adducts. In contrast, the uvrD gene product, which has been found to be involved in the repair of UV damage, has no effect in repairing CC-1065-DNA adducts. Purified UVRA, UVRB, and UVRC proteins must work in concert to incise the drug-modified ΦX174 RFI DNA. Using a site-directed and multiple CC-1065 modified (MspI-BstNI) 117 base pair fragment from M13mpl, we have found that UVRABC nuclease incises at the eighth phosphodiester bond on the 5′ side of the CC-1065-DNA adduct on the drug-modified strand. The enzymes do not cut the noncovalently modified strand. At low drug binding ratios, of the four CC-1065 binding sites identified in the (MspI-BstNI) 117 base pair fragment, GATTA*, GGAAA*, GATAA*, and TTTTA* (* indicates the covalently modified adenine), only the adduct at the high-affinity binding site, GATTA*, is incised by the UVRABC nucleases. No difference in the effect of CC-1065 on local DNA structure, as determined by the DNase I cleavage pattern, was evident among these sites. At high drug binding ratios, a fifth drug binding site, AGCTA*, is identified. At this concentration UVRABC nucleases are unable to incise any of these five CC-1065-DNA adducts. The DNA sequence and/or helix-stabilizing effect of multiple adducts may determine the recognition and/or incision of the drug-DNA adduct by UVRABC nuclease. These results are discussed in relation to the structure of the CC-1065-DNA adduct and the effect of drug binding on local DNA structure.

AB - The recognition and repair of the helix-stabilizing and relatively nondistortive CC-1065-(N3-adenine)-DNA adduct by UVRABC nuclease has been investigated both in vivo with ΦX174 RFI DNA by a transfection assay and in vitro by a site-directed adduct in a 117 base pair fragment from M13mpl. CC-1065 is a potent antitumor antibiotic produced by Streptomyces zelensis which binds within the minor groove of DNA through N3 of adenine. In contrast to the helix-destabilizing and distortive modifications of DNA caused by ultraviolet light or N-acetoxy-2-(acetylamino)fluorene, CC-1065 increases the melting point of DNA and decreases the S1 nuclease activity. Using a viral DNA-Escherichia coli transfection system, we have found that the uvrA, uvrB, and uvrC genes, which code for the major excision repair proteins for UV- and NAAAF-induced DNA damage, are also involved in the repair of CC-1065-DNA adducts. In contrast, the uvrD gene product, which has been found to be involved in the repair of UV damage, has no effect in repairing CC-1065-DNA adducts. Purified UVRA, UVRB, and UVRC proteins must work in concert to incise the drug-modified ΦX174 RFI DNA. Using a site-directed and multiple CC-1065 modified (MspI-BstNI) 117 base pair fragment from M13mpl, we have found that UVRABC nuclease incises at the eighth phosphodiester bond on the 5′ side of the CC-1065-DNA adduct on the drug-modified strand. The enzymes do not cut the noncovalently modified strand. At low drug binding ratios, of the four CC-1065 binding sites identified in the (MspI-BstNI) 117 base pair fragment, GATTA*, GGAAA*, GATAA*, and TTTTA* (* indicates the covalently modified adenine), only the adduct at the high-affinity binding site, GATTA*, is incised by the UVRABC nucleases. No difference in the effect of CC-1065 on local DNA structure, as determined by the DNase I cleavage pattern, was evident among these sites. At high drug binding ratios, a fifth drug binding site, AGCTA*, is identified. At this concentration UVRABC nucleases are unable to incise any of these five CC-1065-DNA adducts. The DNA sequence and/or helix-stabilizing effect of multiple adducts may determine the recognition and/or incision of the drug-DNA adduct by UVRABC nuclease. These results are discussed in relation to the structure of the CC-1065-DNA adduct and the effect of drug binding on local DNA structure.

UR - http://www.scopus.com/inward/record.url?scp=0023846687&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023846687&partnerID=8YFLogxK

M3 - Article

VL - 27

SP - 893

EP - 901

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 3

ER -