Lesion bypass activity of DNA polymerase A from the extremely radioresistant organism Deinococcus radiodurans.
The bacterium Deinococcus radiodurans survives extremely high exposure to ionizing radiation and extended periods of desiccation. Radiation at the survival doses is known to cause numerous DNA damage, such as hundreds of double strand breaks and single strand breaks, as well as damage of the nucleobases. The mechanisms of D. radiodurans to survive the depicted threats are still only beginning to be understood. DNA polymerase A (PolA) has been shown to be crucially involved in irradiation resistance mechanisms of D. radiodurans. We expressed and characterized the DNA polymerase domain of PolA for the first time in vitro. The obtained enzyme is able to efficiently catalyze DNA-dependent DNA synthesis requiring Mg(II) as divalent metal ion. Additionally, strand displacement synthesis of the DNA polymerase, which is required in several repair processes, could be detected. We further found that DNA polymerase function of PolA is modulated by the presence of Mn(II). Whereas proceeding DNA synthesis of PolA was blocked by certain DNA damage that occurs through radiation of DNA, bypass was facilitated by Mn(II). Our results suggest an enzyme modulator function of Mn(II). These observations parallel reports that D. radiodurans accumulates intracellular Mn(II) in cases of irradiation and that the level of irradiation protection correlates with Mn(II) concentrations.
Heinz
Kathrin
K
Department of Chemistry, University of Konstanz, 78457 Konstanz, Germany.
Marx
Andreas
A
D003001:Cloning, Molecular; D004249:DNA Damage / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D034301:Deinococcus / Q000201:enzymology* / Q000235:genetics / Q000528:radiation effects*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic; D007700:Kinetics; D009711:Nucleotides / Q000378:metabolism; D011836:Radiation Tolerance / Q000235:genetics*; D013379:Substrate Specificity
D004269:DNA, Bacterial; D009711:Nucleotides; D004259:DNA-Directed DNA Polymerase