In vitro activities of the multifunctional RNA silencing polymerase QDE-1 of Neurospora crassa.

The Journal of Biological Chemistry
Antti P AaltoDennis H Bamford

Abstract

QDE-1 is an RNA- and DNA-dependent RNA polymerase that has functions in the RNA silencing and DNA repair pathways of the filamentous fungus Neurospora crassa. The crystal structure of the dimeric enzyme has been solved, and the fold of its catalytic core is related closely to that of eukaryotic DNA-dependent RNA polymerases. However, the specific activities of this multifunctional enzyme are still largely unknown. In this study, we characterized the in vitro activities of the N-terminally truncated QDE-1ΔN utilizing structure-based mutagenesis. Our results indicate that QDE-1 displays five distinct catalytic activities, which can be dissected by mutating critical amino acids or by altering reaction conditions. Our data also suggest that the RNA- and DNA-dependent activities have different modes for the initiation of RNA synthesis, which may reflect the mechanism that enables the polymerase to discriminate between template nucleic acids. Moreover, we show that QDE-1 is a highly potent terminal nucleotidyltransferase. Our results suggest that QDE-1 is able to regulate its activity mode depending on the template nucleic acid. This work extends our understanding of the biochemical properties of the QDE-1 enzyme and related RNA poly...Continue Reading

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Citations

Jul 27, 2014·Molecular Biology and Evolution·Heli A M MönttinenMinna M Poranen
Feb 26, 2015·Proceedings of the National Academy of Sciences of the United States of America·Matteo PasettoDavid J FitzGerald
May 1, 2020·Cold Spring Harbor Symposia on Quantitative Biology·Jasleen Singh, Craig S Pikaard
Feb 26, 2016·The Journal of Biological Chemistry·Xinlei QianJulien Lescar
Dec 24, 2018·Biochemical Society Transactions·David Forrest
Nov 20, 2020·Nature·Arina V DrobyshevaMaria L Sokolova
Jan 5, 2018·Bio-protocol·Kyungmin BaegHiro-Oki Iwakawa

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