Comprehensive re-analysis of hairpin small RNAs in fungi reveals loci with conserved links
- Universidad Mayor, Chile
- Pontificia Universidad Católica de Chile, Chile
- Universidad Andrés Bello, Chile
Abstract
RNA interference is an ancient mechanism with many regulatory roles in eukaryotic genomes, with small RNAs acting as their functional element. While there is a wide array of classes of small-RNA-producing loci, those resulting from stem-loop structures (hairpins) have received profuse attention. Such is the case of microRNAs (miRNAs), which have distinct roles in plants and animals. Fungi also produce small RNAs, and several publications have identified miRNAs and miRNA-like (mi/milRNA) hairpin RNAs in diverse fungal species using deep sequencing technologies. Despite this relevant source of information, relatively little is known about mi/milRNA-like features in fungi, mostly due to a lack of established criteria for their annotation. To systematically assess mi/miRNA-like characteristics and annotation confidence, we searched for publications describing mi/milRNA loci and re-assessed the annotations for 41 fungal species. We extracted and normalized the annotation data for 1,727 reported mi/milRNA-like loci and determined their abundance profiles, concluding that less than half of the reported loci passed basic standards used for hairpin RNA discovery. We found that fungal mi/milRNA are generally more similar in size to animal miRNAs and were frequently associated with protein-coding genes. The compiled genomic analyses identified 25 mi/milRNA loci conserved in multiple species. Our pipeline allowed us to build a general hierarchy of locus quality, identifying more than 150 loci with high-quality annotations. We provide a centralized annotation of identified mi/milRNA hairpin RNAs in fungi which will serve as a resource for future research and advance in understanding the characteristics and functions of mi/milRNAs in fungal organisms.
Data availability
Sequencing data used in this work is available in public repositories, with publication details provided in Table S1 and all data accessions provided in Table S3. Results of abundance profiling are found in File S1 and summarized in Table S4.
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Small RNA-seq from multiple Cuscuta species parasitizing ArabidopsisNCBI BioProjects, PRJNA543296.
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Penicillium marneffei Small RNA Transcriptome (17-30nt)NCBI BioProjects, PRJNA207279.
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small RNA and degradome sequencing in Fusarium oxysporumNCBI BioProjects, PRJNA232807.
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Fusarium graminearum strain:HN9-1 Transcriptome or Gene expressionNCBI BioProjects, PRJNA253151.
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Small RNA sequencing of Penicillium chrysogenum P2niaD18NCBI BioProjects, PRJNA270038.
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Taiwanofungus camphoratus Transcriptome or Gene expressionNCBI BioProjects, PRJNA268267.
-
Zymoseptoria tritici small RNA transcriptomeNCBI BioProjects, PRJNA271281.
-
Exploring pathogenic microRNAs of rice sheath blight pathogenNCBI BioProjects, PRJNA282111.
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Puccinia striiformis f. sp. tritici strain:CYR32 Raw sequence readsNCBI BioProjects, PRJNA355964.
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Small RNA sequencing of Coprinopsis cinereaNCBI BioProjects, PRJNA477255.
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Integrated microRNA and mRNA analysis in T. rubrumNCBI BioProjects, PRJNA483837.
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small RNAs sequencing of the Botryosphaeia dothidea strainsNCBI BioProjects, PRJNA511629.
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Sexual development of Cordyceps militarisNCBI BioProjects, PRJNA496418.
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Triticum aestivum cultivar:HD2329 (bread wheat)NCBI BioProjects, PRJNA266709.
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Volvariella volvacea Raw sequence readsNCBI BioProjects, PRJNA594834.
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Interactions between maize and its BLSB pathogenNCBI BioProjects, PRJNA596921.
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Pisolithus microcarpus SI14 smRNA sequencing Redo - SI14_1wk_ECM3JGI GOLD (published), Gp0317539.
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Pisolithus microcarpus SI9 smRNA sequencing Redo - SI9_1wk_ECM1JGI GOLD (published), Gp0317541.
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Pisolithus microcarpus R4 smRNA - R4_FLM3_RedoJGI GOLD (published), Gp0251344.
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Pisolithus microcarpus R10 smRNA - R10_FLM3_RedoJGI GOLD (published), Gp0251333.
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Constructed small RNA libraries from N. bombycisNCBI BioProjects, PRJNA760284.
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small RNA sequences of Fusarium oxysporum f. sp. cubense TR4NCBI BioProjects, PRJNA562097.
Article and author information
Author details
Funding
Fondo Nacional de Desarrollo Científico y Tecnológico (11220727)
- Nathan R Johnson
Instituto Milenio de Biologia Integrativa (ICN17_022)
- Nathan R Johnson
- Luis F Larrondo
- Elena A Vidal
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2022, Johnson et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Comprehensive re-analysis of hairpin small RNAs in fungi reveals loci with conserved links
eLife 11:e83691.
https://doi.org/10.7554/eLife.83691
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Enhancers and promoters are classically considered to be bound by a small set of transcription factors (TFs) in a sequence-specific manner. This assumption has come under increasing skepticism as the datasets of ChIP-seq assays of TFs have expanded. In particular, high-occupancy target (HOT) loci attract hundreds of TFs with often no detectable correlation between ChIP-seq peaks and DNA-binding motif presence. Here, we used a set of 1003 TF ChIP-seq datasets (HepG2, K562, H1) to analyze the patterns of ChIP-seq peak co-occurrence in combination with functional genomics datasets. We identified 43,891 HOT loci forming at the promoter (53%) and enhancer (47%) regions. HOT promoters regulate housekeeping genes, whereas HOT enhancers are involved in tissue-specific process regulation. HOT loci form the foundation of human super-enhancers and evolve under strong negative selection, with some of these loci being located in ultraconserved regions. Sequence-based classification analysis of HOT loci suggested that their formation is driven by the sequence features, and the density of mapped ChIP-seq peaks across TF-bound loci correlates with sequence features and the expression level of flanking genes. Based on the affinities to bind to promoters and enhancers we detected five distinct clusters of TFs that form the core of the HOT loci. We report an abundance of HOT loci in the human genome and a commitment of 51% of all TF ChIP-seq binding events to HOT locus formation thus challenging the classical model of enhancer activity and propose a model of HOT locus formation based on the existence of large transcriptional condensates.
