A novel single-stranded DNA-specific 3′-5′ exonuclease, Thermus thermophilus exonuclease I, is involved in several DNA repair pathways

Abstract

Single-stranded DNA (ssDNA)-specific exonucleases (ssExos) are expected to be involved in a variety of DNA repair pathways corresponding to their cleavage polarities; however, the relationship between the cleavage polarity and the respective DNA repair pathways is only partially understood. To understand the cellular function of ssExos in DNA repair better, genes encoding ssExos were disrupted in Thermus thermophilus HB8 that seems to have only a single set of 5′-3′ and 3′-5′ ssExos unlike other model organisms. Disruption of the tthb178 gene, which was expected to encode a 3′-5′ ssExo, resulted in significant increase in the sensitivity to H2O2 and frequency of the spontaneous mutation rate, but scarcely affected the sensitivity to ultraviolet (UV) irradiation. In contrast, disruption of the recJ gene, which encodes a 5′-3′ ssExo, showed little effect on the sensitivity to H2O2, but caused increased sensitivity to UV irradiation. In vitro characterization revealed that TTHB178 possessed 3′-5′ ssExo activity that degraded ssDNAs containing deaminated and methylated bases, but not those containing oxidized bases or abasic sites. Consequently, we concluded that TTHB178 is a novel 3′-5′ ssExo that functions in various DNA repair systems in cooperation with or independently of RecJ. We named TTHB178 as T. thermophilus exonuclease I. © The Author(s) 2010. Published by Oxford University Press.