Alkaline gel electrophoresis of deoxyribonucleic acid photoreacted with trimethylpsoralen: rapid and sensitive detection of interstrand cross-links

Biochemistry ◽  
1981 ◽  
Vol 20 (6) ◽  
pp. 1431-1437 ◽  
Author(s):  
Thomas R. Cech
Keyword(s):  
Gel Electrophoresis ◽  
Deoxyribonucleic Acid ◽  
Sensitive Detection ◽  
Interstrand Cross Links ◽  
Cross Links

scholarly journals Hydrated electrons induce the formation of interstrand cross-links in DNA modified by cisplatin adducts

2020 ◽  
Vol 61 (3) ◽  
pp. 343-351
Author(s):  
B Behmand ◽  
A M Noronha ◽  
C J Wilds ◽  
J-L Marignier ◽  
M Mostafavi ◽  
...  
Keyword(s):  
Rate Constant ◽  
Gamma Rays ◽  
Gel Electrophoresis ◽  
Pulse Radiolysis ◽  
Interstrand Crosslinks ◽  
Radiosensitizing Effect ◽  
Hydrated Electrons ◽  
Interstrand Cross Links ◽  
Cross Links

Abstract Double-stranded oligonucleotides containing cisplatin adducts, with and without a mismatched region, were exposed to hydrated electrons generated by gamma-rays. Gel electrophoresis analysis demonstrates the formation of cisplatin-interstrand crosslinks from the cisplatin-intrastrand species. The rate constant per base for the reaction between hydrated electrons and the double-stranded oligonucleotides with and without cisplatin containing a mismatched region was determined by pulse radiolysis to be 7 × 109 and 2 × 109 M−1 s−1, respectively. These results provide a better understanding of the radiosensitizing effect of cisplatin adducts in hypoxic tumors and of the formation of interstrand crosslinks, which are difficult for cells to repair.

scholarly journals Chemical and strutural properties of DNA-abasic site cross-links

2014 ◽  
Author(s):  
◽  
Nathan Price
Keyword(s):  
Gel Electrophoresis ◽  
Structural Information ◽  
Sequence Specificity ◽  
Abasic Site ◽  
Cross Link ◽  
Dna Sequence Specificity ◽  
Abasic Sites ◽  
Interstrand Cross Links ◽  
Cross Links ◽  
Synthetic Standard

Here, we report on the formation of DNA interstrand cross-links. These cross-links form from abasic sites, an endogenous type of DNA damage. We have utilized gel electrophoresis, NMR, mass spectrometry and liquid chromatography to identify the location, DNA sequence specificity, stability and some structural information for these interstrand cross-links. Additionally, we have synthesized a small molecule standard which is identical to the enzymatically digested cross-link from duplex DNA. This synthetic standard can be used to increase sensitivity for future detection of DNA-abasic site cross-links.

scholarly journals Optimization of the comet assay for the sensitive detection of PUVA-induced DNA interstrand cross-links

Mutagenesis ◽  
2008 ◽  
Vol 24 (2) ◽  
pp. 173-181 ◽  
Author(s):  
J. H. Wu ◽  
J. B. Wilson ◽  
A. M. Wolfreys ◽  
A. Scott ◽  
N. J. Jones
Keyword(s):  
Comet Assay ◽  
Sensitive Detection ◽  
Interstrand Cross Links ◽  
Cross Links

Electron microscopic study of the membrane glycoproteins in the rat kidney after cisplatin treatment

1989 ◽  
Vol 47 ◽  
pp. 912-913
Author(s):  
S.K. Aggarwal
Keyword(s):  
Alkaline Phosphatase ◽  
Rat Kidney ◽  
Light And Electron Microscopy ◽  
Kidney Tissue ◽  
Membrane Glycoproteins ◽  
Electron Microscopic ◽  
Before And After ◽  
Interstrand Cross Links ◽  
Cross Links

The proposed primary mechanism of action of the anticancer drug cisplatin (Cis-DDP) is through its interaction with DNA, mostly through DNA intrastrand cross-links or DNA interstrand cross-links. DNA repair mechanisms can circumvent this arrest thus permitting replication and transcription to proceed. Various membrane transport enzymes have also been demonstrated to be effected by cisplatin. Glycoprotein alkaline phosphatase was looked at in the proximal tubule cells before and after cisplatin both in vivo and in vitro for its inactivation or its removal from the membrane using light and electron microscopy.Outbred male Swiss Webster (Crl: (WI) BR) rats weighing 150-250g were given ip injections of cisplatin (7mg/kg). Animals were killed on day 3 and day 5. Thick slices (20-50.um) of kidney tissue from treated and untreated animals were fixed in 1% buffered glutaraldehyde and 1% formaldehyde (0.05 M cacodylate buffer, pH 7.3) for 30 min at 4°C. Alkaline phosphatase activity and carbohydrates were demonstrated according to methods described earlier.

Increased gene-specific repair of cisplatin interstrand cross-links in cisplatin-resistant human ovarian cancer cell lines

1992 ◽  
Vol 12 (9) ◽  
pp. 3689-3698
Author(s):  
W Zhen ◽  
C J Link ◽  
P M O'Connor ◽  
E Reed ◽  
R Parker ◽  
...  
Keyword(s):  
Cell Lines ◽  
Ovarian Cancer Cell ◽  
Parental Cell ◽  
Resistant Cell ◽  
Dhfr Gene ◽  
Human Ovarian Cancer Cell ◽  
Ovarian Cancer Cell Lines ◽  
Repair Efficiency ◽  
Interstrand Cross Links ◽  
Cross Links

We have studied several aspects of DNA damage formation and repair in human ovarian cancer cell lines which have become resistant to cisplatin through continued exposure to the anticancer drug. The resistant cell lines A2780/cp70 and 2008/c13*5.25 were compared with their respective parental cell lines, A2780 and 2008. Cells in culture were treated with cisplatin, and the two main DNA lesions formed, intrastrand adducts and interstrand cross-links, were quantitated before and after repair incubation. This quantitation was done for total genomic lesions and at the level of individual genes. In the overall genome, the initial frequency of both cisplatin lesions assayed was higher in the parental than in the derivative resistant cell lines. Nonetheless, the total genomic repair of each of these lesions was not increased in the resistant cells. These differences in initial lesion frequency between parental and resistant cell lines were not observed at the gene level. Resistant and parental cells had similar initial frequencies of intrastrand adducts and interstrand cross-links in the dihydrofolate reductase (DHFR) gene and in several other genes after cisplatin treatment of the cells. There was no increase in the repair efficiency of intrastrand adducts in the DHFR gene in resistant cell lines compared with the parental partners. However, a marked and consistent repair difference between parental and resistant cells was observed for the gene-specific repair of cisplatin interstrand cross-links. DNA interstrand cross-links were removed from three genes, the DHFR, multidrug resistance (MDR1), and delta-globin genes, much more efficiently in the resistant cell lines than in the parental cell lines. Our findings suggest that acquired cellular resistance to cisplatin may be associated with increased gene-specific DNA repair efficiency of a specific lesion, the interstrand cross-link.

scholarly journals The Pso4 mRNA Splicing and DNA Repair Complex Interacts with WRN for Processing of DNA Interstrand Cross-links

2005 ◽  
Vol 280 (49) ◽  
pp. 40559-40567 ◽  
Author(s):  
Nianxiang Zhang ◽  
Ramandeep Kaur ◽  
Xiaoyan Lu ◽  
Xi Shen ◽  
Lei Li ◽  
...  
Keyword(s):  
Dna Repair ◽  
Mrna Splicing ◽  
Interstrand Cross Links ◽  
Cross Links
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