Environmental Temperature Controls Accumulation of Transacting siRNAs Involved in Heterochromatin Formation

Genes
Marcello PirritanoMartin Simon

Abstract

Genes or alleles can interact by small RNAs in a homology dependent manner meaning that short interfering (siRNAs) can act intransat the chromatin level producing stable and heritable silencing phenotypes. Because of the puzzling data on endogenous paramutations, their impact contributing to adaptive evolution in a Lamarckian manner remains unknown. An increasing number of studies characterizes the underlying siRNA accumulation pathways using transgene experiments. Also in the ciliateParamecium tetraurelia, we inducetranssilencing on the chromatin level by injection of truncated transgenes. Here, we characterize the efficiency of this mechanism at different temperatures showing that silencing of the endogenous genes is temperature dependent. Analyzing different transgene constructs at different copy numbers, we dissected whether silencing efficiency is due to varying precursor RNAs or siRNA accumulation. Our data shows that silencing efficiency correlates with more efficient accumulation of primary siRNAs at higher temperatures rather than higher expression of precursor RNAs. Due to higher primary levels, secondary siRNAs also show temperature dependency and interestingly increase their relative proportion to primary siRNAs. Ou...Continue Reading

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Citations

Apr 28, 2020·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Sivarajan KarunanithiMarcel H Schulz
Jun 30, 2019·Nucleic Acids Research·Sivarajan KarunanithiMarcel H Schulz

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Methods Mentioned

BETA
not
Transgenic
electrophoresis
PCR
Illumina Sequencing
RNAseq

Software Mentioned

RAPID
Trim Galore
Cutadapt

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