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Functional analyses of RNA interference effectors in Dictyostelium discoideum during growth and development
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Microbiology. (Fredrik Söderbom)
Uppsala University, Science for Life Laboratory, SciLifeLab. Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Microbiology.
(English)Manuscript (preprint) (Other academic)
Abstract [en]

RNA interference (RNAi) is a widespread biological process, which regulates gene expression in eukaryotic cells. A complex of proteins and small RNAs, RISC, mediates this gene regulation. Central to the function of RISC are the Argonaute effector proteins, which bind the small RNAs, e.g. micro (mi)RNAs and small interfering (si)RNAs. It has previously been shown that RNAi is important to control transposon mobilization in the social amoeba Dictyostelium discoideum, a unicellular eukaryote that upon starvation enters a multicellular developmental program. Information concerning the five Argonautes in D. discoideum is scarce but several of them appear to inhibit transposon mobilization by RNAi related mechanisms. In a recent study, we showed that three of the Argonautes in D. discoideum are involved in controlling cell division. In this study, we perform mRNA- and small RNA-seq. from growing and developing cells, combined with phenotypic studies of D. discoideum strains depleted of AgnB, AgnC, and AgnE. The previously observed effect on cell division, i.e. faster growth for agnE-, and slower for agnB- and agnC- cells, is associated with increased and decreased expression, respectively, of genes involved in nucleotide metabolism. Furthermore, all three argonautes appear to be involved in downregulation of ribosomal protein genes during development while AgnE also contributes to reduced expression of protein coding genes during growth. These effects are likely mediated by small RNAs. We further report the subcellular localization of the three Argonautes, where AgnB is mainly localized in the cytoplasm, AgnC in the nucleus, and the previously reported cytoplasmic localization for AgnE was confirmed. Finally, we present data indicating that AgnB is interacting with miRNAs, suggesting that this Argonaute is involved in miRNA mediated gene regulation in D. discoideum.  Taken together, our data indicate that none of the Argonautes components are essential for cell division and development, but all participate in fine-tuning of gene expression for optimal growth and synchronous multicellular development.

Keywords [en]
RNAi, microRNA, Argonautes, Dictyostelium discoideum, amoeba, development
National Category
Cell Biology Microbiology
Research subject
Biology with specialization in Molecular Cell Biology
Identifiers
URN: urn:nbn:se:uu:diva-354021OAI: oai:DiVA.org:uu-354021DiVA, id: diva2:1221016
Funder
Swedish National Infrastructure for Computing (SNIC), 621-2013-4665Carl Tryggers foundation , CST12:485Available from: 2018-06-19 Created: 2018-06-19 Last updated: 2018-06-19
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