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Excision-Repair of γ-Ray-Damaged Thymine in Bacterial and Mammalian Systems

  • P. V. Hariharan
  • J. F. Remsen
  • P. A. Cerutti
Part of the Basic Life Sciences book series

Abstract

The selective excision of products of the 5,6-dihydroxy-dihydrothymine type (t′) from γ-irradiated or 0s04-oxidized DNA or synthetic poly[d(A-T)] was observed with crude extracts of Escherichia coli and isolated nuclei from human carcinoma HeLa S-3 cells and Chinese hamster ovary cells.

The results with E. coli extracts allow the following conclusion: (1) The uvrA-gene product is not required for t′ excision. (2) Radiation-induced strand breakage is not required for product excision. (3) Experiments with extracts of E. coli polAexl showed that the 5′→3′ exonuclease associated with polymerase I is responsible for the removal of t′. (4) Experiments with extracts of E. coli endo I rig 4 and the ligase inhibitor nicotinamide mononucleotide showed that polynucleotide ligase accomplishes the last strand resealing step in the excision-repair of t′.

Isolated nuclei from HeLa and Chinese hamster ovary cells possess the necessary enzymes for the selective excision of t′ from γ-irradiated or osmium tetroxide oxidized DNA. Approximately 25 to 35% of the products were removed from DNA within 60 min. Unspecific DNA degradation was very low. Radiation-induced strand breakage is not required for product removal.

Keywords

Base Damage Strand Breakage Thymine Residue Repair Replication Product Excision 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1975

Authors and Affiliations

  • P. V. Hariharan
    • 1
  • J. F. Remsen
    • 1
  • P. A. Cerutti
    • 1
  1. 1.Department of BiochemistryUniversity of FloridaGainesvilleUSA

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