Formation and repair of unavoidable, endogenous interstrand cross-links in cellular DNA

Diverse applications of covalent cross-links in duplex DNA: From anti-cancer drugs to the detection of disease-relevant single nucleotide polymorphisms to the synthesis of well-defined substrates for the study of novel DNA repair processes

10.32469/10355/69816 ◽

2018 ◽

Author(s):

◽

Maryam Imani Nejad

Keyword(s):

Excision Repair ◽

Alkylating Agents ◽

Nucleotide Polymorphisms ◽

Cross Link ◽

Single Nucleotide ◽

Synthetic Dna ◽

Interstrand Cross Links ◽

Cross Links ◽

Ap Sites ◽

Cellular Dna

[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] Abasic (Ap) sites are a common form of DNA lesion that occur endogenously 50,000-200,000 per cell per day in mammals. The alkylation of the guanine and adenine residues by the alkylating agents such as nitrogen mustards also induces the formation of Ap sites in genomic DNA. Our group recently showed that Ap sites can forge DNA-DNA interstrand cross-links in some sequences via reaction of the Ap aldehyde residue with the exocyclic amino groups of nucleobases, such as adenine and guanine, on the opposing strand of the DNA duplex. The earlier work in the group revealed that formation of these covalent bridges between two DNA strands is highly sequence- dependent. Although interstrand cross-links are one of the most deleterious types of cellular DNA damage, the availability of synthetic DNA duplexes containing chemically well-defined, site-specific interstrand cross-links has been proven to be a valuable tool in biological chemistry and medicine. We prepared and characterized a new Ap-derived interstrand cross-link. In another project, we use these remarkable cross-linking reactions for the covalent capture of disease-relevant single nucleotide polymorphism by using a protein nanopore technology. The complex mechanisms underlying cross-link repair in cells and limited availability of stable and defined cross-link are two major reasons why repair pathways of these lesions are not yet well understood. By preparing a variety of Ap-derived cross-links, we studied the role of a base excision repair DNA glycosylase, NEIL3 in unhooking the lesions.

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Rescue of mitomycin C- or psoralen-inactivated Micrococcus radiodurans by additional exposure to radiation or alkylating agents

Journal of Bacteriology ◽

10.1128/jb.152.3.976-982.1982 ◽

1982 ◽

Vol 152 (3) ◽

pp. 976-982

Author(s):

M T Hansen

Keyword(s):

Mitomycin C ◽

Uv Radiation ◽

Uv Light ◽

Alkylating Agents ◽

Repair System ◽

Wild Type ◽

Interstrand Cross Links ◽

Cross Links ◽

Cellular Dna ◽

Constitutive Synthesis

The processing of damaged DNA was altered in a mitomycin C-sensitive mutant (mtcA) of Micrococcus radiodurans. Even though the mutant retained resistance to 254-nm UV radiation, it did not, in contrast to the wild-type strain, show any excessive DNA degradation or cell death when incubated with chloramphenicol after sublethal doses of either UV light or mitomycin C. The results suggest the constitutive synthesis of an enzyme system responsible for wild-type proficiency in the repair of mitomycin C-induced damage. An alternative system able to repair damage caused by mitomycin C was demonstrated in the mtcA background. In this strain, additional damage inflicted upon the cellular DNA effected a massive rescue of cells previously inactivated by mitomycin C. Rescue was provoked by ionizing radiation, by UV light, or by simple alkylating agents. Cells treated with psoralen plus near-UV radiation could be rescued only when inactivation was due primarily to psoralen-DNA interstrand cross-links rather than to monoadducts. The rescue of inactivated cells was prevented in the presence of chloramphenicol. These results can be interpreted most readily in terms of an alternative repair system able to overcome DNA interstrand cross-links produced by mitomycin C or psoralen plus near-UV light, but induced only by the more abundant number of damages produced by radiation or simple alkylating agents.

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

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100156547 ◽

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.

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Correction to Nucleotide Excision Repair of Chemically Stabilized Analogues of DNA Interstrand Cross-Links Produced from Oxidized Abasic Sites

Biochemistry ◽

10.1021/bi501162v ◽

2014 ◽

Vol 53 (40) ◽

pp. 6418-6418

Author(s):

Souradyuti Ghosh ◽

Marc M. Greenberg

Keyword(s):

Nucleotide Excision Repair ◽

Excision Repair ◽

Abasic Sites ◽

Nucleotide Excision ◽

Interstrand Cross Links ◽

Cross Links

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DNA interstrand cross-links from modified nucleotides: mechanism and application

Nucleic Acids Symposium Series ◽

10.1093/nass/49.1.57 ◽

2005 ◽

Vol 49 (1) ◽

pp. 57-58 ◽

Cited By ~ 3

Author(s):

Marc M. Greenberg

Keyword(s):

Modified Nucleotides ◽

Interstrand Cross Links ◽

Cross Links

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

Molecular and Cellular Biology ◽

10.1128/mcb.12.9.3689-3698.1992 ◽

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.

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