Structure of the DNA Repair Helicase Hel308 Reveals DNA Binding and Autoinhibitory Domains

• Published in: The Journal of biological chemistry Vol. 283; no. 8; pp. 5118 - 5126
• Main Authors: Richards, Jodi D., Johnson, Kenneth A., Liu, Huanting, McRobbie, Anne-Marie, McMahon, Stephen, Oke, Muse, Carter, Lester, Naismith, James H., White, Malcolm F.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 22.02.2008; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; Animals; Archaea; Archaeal Proteins - chemistry; Archaeal Proteins - genetics; Archaeal Proteins - metabolism; Crystallography, X-Ray; DNA Helicases - chemistry; DNA Helicases - genetics; DNA Helicases - metabolism; DNA Repair - physiology; DNA, Archaeal - chemistry; DNA, Archaeal - genetics; DNA, Archaeal - metabolism; DNA, Single-Stranded - chemistry; DNA, Single-Stranded - genetics; DNA, Single-Stranded - metabolism; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; DNA-Directed DNA Polymerase - chemistry; DNA-Directed DNA Polymerase - genetics; DNA-Directed DNA Polymerase - metabolism; Drosophila; Humans; Molecular Sequence Data; Protein Binding - physiology; Protein Structure, Tertiary - physiology; Recombination, Genetic - physiology; Structural Homology, Protein; Sulfolobus solfataricus; Sulfolobus solfataricus - enzymology; Sulfolobus solfataricus - genetics
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M707548200

Hel308 is a superfamily 2 helicase conserved in eukaryotes and archaea. It is thought to function in the early stages of recombination following replication fork arrest and has a specificity for removal of the lagging strand in model replication forks. A homologous helicase constitutes the N-terminal domain of human DNA polymerase Q. The Drosophila homologue mus301 is implicated in double strand break repair and meiotic recombination. We have solved the high resolution crystal structure of Hel308 from the crenarchaeon Sulfolobus solfataricus, revealing a five-domain structure with a central pore lined with essential DNA binding residues. The fifth domain is shown to act as an autoinhibitory domain or molecular brake, clamping the single-stranded DNA extruded through the central pore of the helicase structure to limit the helicase activity of the enzyme. This provides an elegant mechanism to tune the processivity of the enzyme to its functional role. Hel308 can displace streptavidin from a biotinylated DNA molecule, and this activity is only partially inhibited when the DNA is pre-bound with abundant DNA-binding proteins RPA or Alba1, whereas pre-binding with the recombinase RadA has no effect on activity. These data suggest that one function of the enzyme may be in the removal of bound proteins at stalled replication forks and recombination intermediates.