Biochemical characterization of AtRECQ3 reveals significant differences relative to other RecQ helicases.
Members of the conserved RecQ helicase family are important for the preservation of genomic stability. Multiple RecQ homologs within one organism raise the question of functional specialization. Whereas five different homologs are present in humans, the model plant Arabidopsis (Arabidopsis thaliana) carries seven RecQ homologs in its genome. We performed biochemical analysis of AtRECQ3, expanded upon a previous analysis of AtRECQ2, and compared their properties. Both proteins differ in their domain composition. Our analysis demonstrates that they are 3' to 5' helicases with similar activities on partial duplex DNA. However, they promote different outcomes with synthetic DNA structures that mimic Holliday junctions or a replication fork. AtRECQ2 catalyzes Holliday junction branch migration and replication fork regression, while AtRECQ3 cannot act on intact Holliday junctions. The observed reaction of AtRECQ3 on the replication fork is in line with unwinding the lagging strand. On nicked Holliday junctions, which have not been intensively studied with RecQ helicases before, AtRECQ3, but not AtRECQ2, shows a clear preference for one unwinding mechanism. In addition, AtRECQ3 is much more efficient at catalyzing DNA strand annealing. Thus, AtRECQ2 and AtRECQ3 are likely to perform different tasks in the cell, and AtRECQ3 differs in its biochemical properties from all other eukaryotic RECQ helicases characterized so far.
Kobbe
Daniela
D
Botany II, University of Karlsruhe, 76128 Karlsruhe, Germany.
Blanck
Sandra
S
Focke
Manfred
M
Puchta
Holger
H
D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D004261:DNA Replication; D045566:DNA, Cruciform / Q000378:metabolism*; D018744:DNA, Plant / Q000378:metabolism*; D003854:Deoxyribonucleotides / Q000378:metabolism; D008969:Molecular Sequence Data; D053484:RecQ Helicases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D045566:DNA, Cruciform; D018744:DNA, Plant; D003854:Deoxyribonucleotides; D053484:RecQ Helicases