Repair of oligodeoxynucleotides containing o-methylguanine by o-alkylguanine-dna-alkyltransferase

David Scicchitano, Roger A. Jones, Sandra Kuzmich, Barbara Gaffney, Dana D. Lasko, John M. Essigmann, Anthony E. Pegg

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

O6-Alkylguanine-DNA-alkyltransferase is a DNA repair protein known to carry out the transfer of alkyl groups from the O6-position of guanine in alkylated DNA to a cysteine acceptor site contained within its own protein sequence. We have examined the ability of this protein isolated from either E. coli or mammalian cells to perform this repair reaction in short oligodeoxynucleotides. Dodecadeoxynucleotides of the sequence 5'-dCGNGAATTCm6GCG-3' where N is any one of the normal four bases were all repaired very rapidly by the protein with 50% repair in < 15 s at 0°C. The hexadeoxy-nucleotide 5'-dCGCm6GCG-3' was repaired slightly more slowly with 50% removal taking 7 min at 0°C and 1.5 min at 37°C. The tetradeoxynucleotide 5'-dTm6GCA-3' was also a substrate but was repaired much more slowly requiring 45 min for 50% repair at 37°C. These results indicate that (a) the AGT has a strong but not absolute preference for double-stranded DNA substrates; (b) the repair of O6-methylguanine is independent of the base opposite the lesion; and (c) that oligodeoxynucleotides as short as tetramers are substrates for repair by this protein.

Original languageEnglish (US)
Pages (from-to)1383-1386
Number of pages4
JournalCarcinogenesis
Volume7
Issue number8
DOIs
StatePublished - Dec 1 1986

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Alkyl and Aryl Transferases
Oligodeoxyribonucleotides
Proteins
DNA
Guanine
DNA Repair
Cysteine
Nucleotides
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Cancer Research

Cite this

Scicchitano, D., Jones, R. A., Kuzmich, S., Gaffney, B., Lasko, D. D., Essigmann, J. M., & Pegg, A. E. (1986). Repair of oligodeoxynucleotides containing o-methylguanine by o-alkylguanine-dna-alkyltransferase. Carcinogenesis, 7(8), 1383-1386. https://doi.org/10.1093/carcin/7.8.1383
Scicchitano, David ; Jones, Roger A. ; Kuzmich, Sandra ; Gaffney, Barbara ; Lasko, Dana D. ; Essigmann, John M. ; Pegg, Anthony E. / Repair of oligodeoxynucleotides containing o-methylguanine by o-alkylguanine-dna-alkyltransferase. In: Carcinogenesis. 1986 ; Vol. 7, No. 8. pp. 1383-1386.
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abstract = "O6-Alkylguanine-DNA-alkyltransferase is a DNA repair protein known to carry out the transfer of alkyl groups from the O6-position of guanine in alkylated DNA to a cysteine acceptor site contained within its own protein sequence. We have examined the ability of this protein isolated from either E. coli or mammalian cells to perform this repair reaction in short oligodeoxynucleotides. Dodecadeoxynucleotides of the sequence 5'-dCGNGAATTCm6GCG-3' where N is any one of the normal four bases were all repaired very rapidly by the protein with 50{\%} repair in < 15 s at 0°C. The hexadeoxy-nucleotide 5'-dCGCm6GCG-3' was repaired slightly more slowly with 50{\%} removal taking 7 min at 0°C and 1.5 min at 37°C. The tetradeoxynucleotide 5'-dTm6GCA-3' was also a substrate but was repaired much more slowly requiring 45 min for 50{\%} repair at 37°C. These results indicate that (a) the AGT has a strong but not absolute preference for double-stranded DNA substrates; (b) the repair of O6-methylguanine is independent of the base opposite the lesion; and (c) that oligodeoxynucleotides as short as tetramers are substrates for repair by this protein.",
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Scicchitano, D, Jones, RA, Kuzmich, S, Gaffney, B, Lasko, DD, Essigmann, JM & Pegg, AE 1986, 'Repair of oligodeoxynucleotides containing o-methylguanine by o-alkylguanine-dna-alkyltransferase', Carcinogenesis, vol. 7, no. 8, pp. 1383-1386. https://doi.org/10.1093/carcin/7.8.1383

Repair of oligodeoxynucleotides containing o-methylguanine by o-alkylguanine-dna-alkyltransferase. / Scicchitano, David; Jones, Roger A.; Kuzmich, Sandra; Gaffney, Barbara; Lasko, Dana D.; Essigmann, John M.; Pegg, Anthony E.

In: Carcinogenesis, Vol. 7, No. 8, 01.12.1986, p. 1383-1386.

Research output: Contribution to journalArticle

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T1 - Repair of oligodeoxynucleotides containing o-methylguanine by o-alkylguanine-dna-alkyltransferase

AU - Scicchitano, David

AU - Jones, Roger A.

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AU - Lasko, Dana D.

AU - Essigmann, John M.

AU - Pegg, Anthony E.

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N2 - O6-Alkylguanine-DNA-alkyltransferase is a DNA repair protein known to carry out the transfer of alkyl groups from the O6-position of guanine in alkylated DNA to a cysteine acceptor site contained within its own protein sequence. We have examined the ability of this protein isolated from either E. coli or mammalian cells to perform this repair reaction in short oligodeoxynucleotides. Dodecadeoxynucleotides of the sequence 5'-dCGNGAATTCm6GCG-3' where N is any one of the normal four bases were all repaired very rapidly by the protein with 50% repair in < 15 s at 0°C. The hexadeoxy-nucleotide 5'-dCGCm6GCG-3' was repaired slightly more slowly with 50% removal taking 7 min at 0°C and 1.5 min at 37°C. The tetradeoxynucleotide 5'-dTm6GCA-3' was also a substrate but was repaired much more slowly requiring 45 min for 50% repair at 37°C. These results indicate that (a) the AGT has a strong but not absolute preference for double-stranded DNA substrates; (b) the repair of O6-methylguanine is independent of the base opposite the lesion; and (c) that oligodeoxynucleotides as short as tetramers are substrates for repair by this protein.

AB - O6-Alkylguanine-DNA-alkyltransferase is a DNA repair protein known to carry out the transfer of alkyl groups from the O6-position of guanine in alkylated DNA to a cysteine acceptor site contained within its own protein sequence. We have examined the ability of this protein isolated from either E. coli or mammalian cells to perform this repair reaction in short oligodeoxynucleotides. Dodecadeoxynucleotides of the sequence 5'-dCGNGAATTCm6GCG-3' where N is any one of the normal four bases were all repaired very rapidly by the protein with 50% repair in < 15 s at 0°C. The hexadeoxy-nucleotide 5'-dCGCm6GCG-3' was repaired slightly more slowly with 50% removal taking 7 min at 0°C and 1.5 min at 37°C. The tetradeoxynucleotide 5'-dTm6GCA-3' was also a substrate but was repaired much more slowly requiring 45 min for 50% repair at 37°C. These results indicate that (a) the AGT has a strong but not absolute preference for double-stranded DNA substrates; (b) the repair of O6-methylguanine is independent of the base opposite the lesion; and (c) that oligodeoxynucleotides as short as tetramers are substrates for repair by this protein.

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Scicchitano D, Jones RA, Kuzmich S, Gaffney B, Lasko DD, Essigmann JM et al. Repair of oligodeoxynucleotides containing o-methylguanine by o-alkylguanine-dna-alkyltransferase. Carcinogenesis. 1986 Dec 1;7(8):1383-1386. https://doi.org/10.1093/carcin/7.8.1383