Topoisomerase IIβ regulates base excision repair capacity of neurons

Abstract

Topoisomerase IIβ (TopoIIβ), an enzyme involved in DNA rearrangements, is predominantly present in brain and its levels are shown to decrease with age. This study characterizes the function of TopoIIβ in regulating BER (base excision repair) activity. TopoIIβ deficient granule neurons (CGNT⁻) show greater sensitivity to N-ethyl N-nitroso urea (ENU)-mediated DNA damage. The cell-free extracts of TopoIIβ knockdown cells (ECGNT⁻) show a significant decrease in G-U BER activity during ENU-treatment as well as during recovery, suggesting that TopoIIβ promotes G-U BER activity. Since G-U BER activity is not affected in the presence of ICRF-193, catalytic inhibitor of TopoIIβ, the activity of enzyme per se may not be participating in BER activity. Further characterization of the activities of BER enzymes present in ECGNT⁻ shows that uracil DNA-glycosylase (UDG) and ligase (LIG) activities decrease significantly in both ENU treatment and recovery. Supplementation of TopoIIβ to ECGNT⁻ does not restore ligation activity and ICRF-193 does not influence the LIG activity. These results suggest a role, at least an indirect one, of TopoIIβ in the repair of ENU-mediated strand breaks via BER pathway including the activities of UDG and LIG.