The Structural and Biochemical Characterization of Human RNase H2 Complex Reveals the Molecular Basis for Substrate Recognition and Aicardi-Goutières Syndrome Defects

Laboratory of Protein Structure, International Institute of Molecular and Cell Biology, Warsaw 02-109, Poland.
Journal of Biological Chemistry (Impact Factor: 4.57). 12/2010; 286(12):10540-50. DOI: 10.1074/jbc.M110.181974
Source: PubMed


RNase H2 cleaves RNA sequences that are part of RNA/DNA hybrids or that are incorporated into DNA, thus, preventing genomic instability and the accumulation of aberrant nucleic acid, which in humans induces Aicardi-Goutières syndrome, a severe autoimmune disorder. The 3.1 Å crystal structure of human RNase H2 presented here allowed us to map the positions of all 29 mutations found in Aicardi-Goutières syndrome patients, several of which were not visible in the previously reported mouse RNase H2. We propose the possible effects of these mutations on the protein stability and function. Bacterial and eukaryotic RNases H2 differ in composition and substrate specificity. Bacterial RNases H2 are monomeric proteins and homologs of the eukaryotic RNases H2 catalytic subunit, which in addition possesses two accessory proteins. The eukaryotic RNase H2 heterotrimeric complex recognizes RNA/DNA hybrids and (5')RNA-DNA(3')/DNA junction hybrids as substrates with similar efficiency, whereas bacterial RNases H2 are highly specialized in the recognition of the (5')RNA-DNA(3') junction and very poorly cleave RNA/DNA hybrids in the presence of Mg(2+) ions. Using the crystal structure of the Thermotoga maritima RNase H2-substrate complex, we modeled the human RNase H2-substrate complex and verified the model by mutational analysis. Our model indicates that the difference in substrate preference stems from the different position of the crucial tyrosine residue involved in substrate binding and recognition.

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    • "Furthermore, both Arg186 and Arg235 are thought to have roles in stabilizing the conformation of a highly conserved GRG motif (Supp. Figure S1B and C) responsible for recognition of the substrate RNA-DNA junction (Figiel, et al., 2010), likely explaining why the Arg186Trp and Arg235Gln mutations dramatically reduce substrate binding and enzyme activity in vitro (Coffin, et al., 2011). The Asn212Ile substitution has no detectable effect on the catalytic activity of recombinant RNase H2, although it does cause a small reduction in substrate binding affinity (Coffin, et al., 2011). "
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