Distinct roles of Argonaute in the green alga Chlamydomonas reveal evolutionary conserved mode of miRNA-mediated gene expression

Scientific Reports
Betty Y ChungDavid C Baulcombe

Abstract

The unicellular green alga Chlamydomonas reinhardtii is evolutionarily divergent from higher plants, but has a fully functional silencing machinery including microRNA (miRNA)-mediated translation repression and mRNA turnover. However, distinct from the metazoan machinery, repression of gene expression is primarily associated with target sites within coding sequences instead of 3'UTRs. This feature indicates that the miRNA-Argonaute (AGO) machinery is ancient and the primary function is for post transcriptional gene repression and intermediate between the mechanisms in the rest of the plant and animal kingdoms. Here, we characterize AGO2 and 3 in Chlamydomonas, and show that cytoplasmically enriched Cr-AGO3 is responsible for endogenous miRNA-mediated gene repression. Under steady state, mid-log phase conditions, Cr-AGO3 binds predominantly miR-C89, which we previously identified as the predominant miRNA with effects on both translation repression and mRNA turnover. In contrast, the paralogue Cr-AGO2 is nuclear enriched and exclusively binds to 21-nt siRNAs. Further analysis of the highly similar Cr-AGO2 and Cr-AGO 3 sequences (90% amino acid identity) revealed a glycine-arginine rich N-terminal extension of ~100 amino acids tha...Continue Reading

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Citations

Nov 20, 2020·PloS One·Sebastian Y MüllerDavid C Baulcombe
Apr 25, 2021·Journal of Experimental Botany·Indu SanthanagopalanHoward Griffiths

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Methods Mentioned

BETA
immunoprecipitation
RNA-seq
PCR
smallRNA-Seq
immunoprecipitations
reverse-phase chromatography

Software Mentioned

R
segmentSeq
Mascot
shell
Protein Discoverer

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