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Oxidant-induced restriction polymorphism maps to kinase region of c-abl oncogene
1Department of Medicine, Northwestern University Medical School, Chicago, Illinois 60611.
Abstract:
Reactive oxygen species generated by activated human phagocytes can cause a variety of genetic injuries and produce malignant transformation in target cells. We previously reported that DNA extracted from phagocyte-transformed 10T1/2 mouse fibroblasts contained Msp I-dependent restriction fragment length polymorphisms in the c-abl oncogene. The data suggested that the oxidant-induced RFLP resulted from an alteration in the methylation pattern in c-abl. We have now mapped one of these RFLP to a specific 'CCGG' tetramer found within the tyrosine kinase region of the gene. The polymorphic 'CCGG' site has been localized to the intron between exon 2 and 3a. Restriction analysis indicates that a repetitive sequence exists within this intron and that the RFLP is associated with this repeat.
Insights
Reactive oxygen species from phagocytes can damage DNA, leading to cancer. This study identifies a specific DNA methylation change in the c-abl oncogene linked to this damage.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- Activated human phagocytes generate reactive oxygen species (ROS).
- ROS can cause genetic damage and malignant transformation in cells.
- Previous work showed ROS-induced DNA alterations in the c-abl oncogene.
Purpose of the Study:
- To map the specific location of oxidant-induced restriction fragment length polymorphisms (RFLPs) in the c-abl oncogene.
- To investigate the association between RFLPs and DNA methylation patterns.
Main Methods:
- DNA extraction from phagocyte-transformed mouse fibroblasts.
- Restriction fragment length polymorphism (RFLP) analysis.
- DNA methylation pattern analysis.
Main Results:
- A specific Msp I-dependent RFLP was mapped to a 'CCGG' tetramer within the c-abl oncogene's tyrosine kinase region.
- The polymorphic 'CCGG' site is located in the intron between exon 2 and 3a.
- Restriction analysis revealed a repetitive sequence in this intron associated with the RFLP.
Conclusions:
- Oxidant-induced RFLPs in the c-abl oncogene are linked to alterations in DNA methylation patterns.
- The identified RFLP is associated with a repetitive sequence in an intron of the c-abl gene.
- These findings contribute to understanding the genetic mechanisms of ROS-induced cell transformation.