A fungal Argonaute interferes with RNA interference

Nucleic Acids Research
Quyet NguyenHitoshi Nakayashiki

Abstract

Small RNA (sRNA)-mediated gene silencing phenomena, exemplified by RNA interference (RNAi), require a unique class of proteins called Argonautes (AGOs). An AGO protein typically forms a protein-sRNA complex that contributes to gene silencing using the loaded sRNA as a specificity determinant. Here, we show that MoAGO2, one of the three AGO genes in the fungus Pyricularia oryzae (Magnaporthe oryzae) interferes with RNAi. Gene knockout (KO) studies revealed that MoAGO1 and MoAGO3 additively or redundantly played roles in hairpin RNA- and retrotransposon (MAGGY)-triggered RNAi while, surprisingly, the KO mutants of MoAGO2 (Δmoago2) showed elevated levels of gene silencing. Consistently, transcript levels of MAGGY and mycoviruses were drastically reduced in Δmoago2, supporting the idea that MoAGO2 impeded RNAi against the parasitic elements. Deep sequencing analysis revealed that repeat- and mycovirus-derived small interfering RNAs were mainly associated with MoAGO2 and MoAGO3, and their populations were very similar based on their size distribution patterns and positional base preference. Site-directed mutagenesis studies indicated that sRNA binding but not slicer activity of MoAGO2 was essential for the ability to diminish the ef...Continue Reading

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Citations

Aug 10, 2019·Phytopathology·Maria D D García-PedrajasJosé Sebastián Dambolena
Apr 2, 2020·International Journal of Molecular Sciences·Ana Lúcia LeitãoFrancisco J Enguita
Jun 12, 2019·International Journal of Molecular Sciences·Chen ZhuCheng-Guo Duan
Nov 20, 2019·Interdisciplinary Sciences, Computational Life Sciences·Lixu JiangJian Huang
Jun 2, 2018·Current Opinion in Plant Biology·Zeyang Ma, Xiuren Zhang
Oct 22, 2018·Fungal Biology·Stefan KuschRalph Panstruga

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Datasets Mentioned

BETA
EHA21073
ETS02811
DRA005932

Methods Mentioned

BETA
gene knockout
PCR
electrophoresis
RNA-Seq
AGO KO
immunoprecipitation
FLAG-immunoprecipitation
RIP

Software Mentioned

DICER
9000
InterPro
Genomics workbench
BZ
KEYENCE
InterPro Scan
FLUOVIEW FV1000 MPE

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