Abstract
The sequence specificity of bleomycin damage was investigated utilising 340 bp alpha-DNA (a middle repetitive sequence in the human genome) as a target sequence. The following significant facts were found:- i) The dinucleotides GT and GC were cleaved on all occasions, GA most of the time, and AT, AC, GG and AA cleaved some of the time; ii) The base immediately 5' to the purine-pyrimidine dinucleotides was found to be statistically highly significant in determining the degree of damage caused by bleomycin, while other nearest neighbour bases had no significant effect; iii) The sequence specificity of bleomycin damage was determined on both strands and it was found that damage on either strand follows the above dinucleotide preference and is independent of the extent of damage on the opposite strand; iv) Bleomycin damage was compared between genomic 340 bp alpha-DNA and a cloned alpha-DNA with eleven base substitutions relative to the "consensus" sequence. There were forty-nine detectable differences in intensity of damage between these two DNA molecules. Although four of the differences can be directly attributed to changes in base sequence, the remaining differences were not at the base substitution sites. Some of the differences were over fifty base pairs from the nearest base substitution. We propose that the majority of these differences are due to microvariation in the structure of DNA with a slightly different DNA sequence.
Full text
PDF














Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Blum R. H., Carter S. K., Agre K. A clinical review of bleomycin--a new antineoplastic agent. Cancer. 1973 Apr;31(4):903–914. doi: 10.1002/1097-0142(197304)31:4<903::aid-cncr2820310422>3.0.co;2-n. [DOI] [PubMed] [Google Scholar]
- Burger R. M., Berkowitz A. R., Peisach J., Horwitz S. B. Origin of malondialdehyde from DNA degraded by Fe(II) x bleomycin. J Biol Chem. 1980 Dec 25;255(24):11832–11838. [PubMed] [Google Scholar]
- Burger R. M., Horwitz S. B., Peisach J., Wittenberg J. B. Oxygenated iron bleomycin. A short-lived intermediate in the reaction of ferrous bleomycin with O2. J Biol Chem. 1979 Dec 25;254(24):12999–12302. [PubMed] [Google Scholar]
- D'Andrea A. D., Haseltine W. A. Sequence specific cleavage of DNA by the antitumor antibiotics neocarzinostatin and bleomycin. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3608–3612. doi: 10.1073/pnas.75.8.3608. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Darling S. M., Crampton J. M., Williamson R. Organization of a family of highly repetitive sequences within the human genome. J Mol Biol. 1982 Jan 5;154(1):51–63. doi: 10.1016/0022-2836(82)90416-8. [DOI] [PubMed] [Google Scholar]
- Dickerson R. E., Drew H. R. Structure of a B-DNA dodecamer. II. Influence of base sequence on helix structure. J Mol Biol. 1981 Jul 15;149(4):761–786. doi: 10.1016/0022-2836(81)90357-0. [DOI] [PubMed] [Google Scholar]
- Dretzen G., Bellard M., Sassone-Corsi P., Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981 Apr;112(2):295–298. doi: 10.1016/0003-2697(81)90296-7. [DOI] [PubMed] [Google Scholar]
- Drew H. R., Wing R. M., Takano T., Broka C., Tanaka S., Itakura K., Dickerson R. E. Structure of a B-DNA dodecamer: conformation and dynamics. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2179–2183. doi: 10.1073/pnas.78.4.2179. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grunberg S. M., Haseltine W. A. Use of an indicator sequence of human DNA to study DNA damage by methylbis(2-chloroethyl)amine. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6546–6550. doi: 10.1073/pnas.77.11.6546. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kross J., Henner W. D., Hecht S. M., Haseltine W. A. Specificity of deoxyribonucleic acid cleavage by bleomycin, phleomycin, and tallysomycin. Biochemistry. 1982 Aug 31;21(18):4310–4318. doi: 10.1021/bi00261a021. [DOI] [PubMed] [Google Scholar]
- Lippke J. A., Gordon L. K., Brash D. E., Haseltine W. A. Distribution of UV light-induced damage in a defined sequence of human DNA: detection of alkaline-sensitive lesions at pyrimidine nucleoside-cytidine sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3388–3392. doi: 10.1073/pnas.78.6.3388. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lown J. W., Sim S. K. The mechanism of the bleomycin-induced cleavage of DNA. Biochem Biophys Res Commun. 1977 Aug 22;77(4):1150–1157. doi: 10.1016/s0006-291x(77)80099-5. [DOI] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Nordheim A., Rich A. Negatively supercoiled simian virus 40 DNA contains Z-DNA segments within transcriptional enhancer sequences. Nature. 1983 Jun 23;303(5919):674–679. doi: 10.1038/303674a0. [DOI] [PubMed] [Google Scholar]
- Takeshita M., Grollman A. P., Ohtsubo E., Ohtsubo H. Interaction of bleomycin with DNA. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5983–5987. doi: 10.1073/pnas.75.12.5983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thayer R. E., Singer M. F., McCutchan T. F. Sequence relationships between single repeat units of highly reiterated African Green monkey DNA. Nucleic Acids Res. 1981 Jan 10;9(1):169–181. doi: 10.1093/nar/9.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu J. C., Manuelidis L. Sequence definition and organization of a human repeated DNA. J Mol Biol. 1980 Sep 25;142(3):363–386. doi: 10.1016/0022-2836(80)90277-6. [DOI] [PubMed] [Google Scholar]


