E. coli DNA Repair Helicase Lhr is also a Uracil-DNA Glycosylase
Abstract
DNA glycosylases protect genetic fidelity during DNA replication by
removing potentially mutagenic chemically damaged DNA bases. Bacterial
Lhr proteins are well-characterized DNA repair helicases that are fused
to additional 600-700 amino acids of unknown function, but with
structural homology to SecB chaperones and AlkZ DNA glycosylases. Here
we identify that E. coli Lhr is a uracil-DNA glycosylase that
depends on an active site aspartic acid residue. We show that the Lhr
DNA helicase activity is functionally independent of the uracil-DNA
glycosylase activity, but that the helicase domains are required for
fully active uracil DNA glycosylase activity. Consistent with uracil DNA
glycosylase activity, deletion of lhr from the E. coli
chromosome sensitized cells to oxidative stress that triggers cytosine
deamination to uracil. The ability of Lhr to translocate single-stranded
DNA and remove uracil bases suggests a surveillance role to seek and
remove potentially mutagenic base changes during replication stress.