Noncoding RNAs (ncRNAs) play important regulatory and functional roles in microorganisms, such as regulation of gene expression, signaling, protein synthesis, and RNA processing. Hence, their classification and quantification are central tasks toward the understanding of the function of the microbial community. However, the majority of the current metagenomic sequencing technologies generate short reads, which may contain only a partial secondary structure that complicates ncRNA homology detection. Meanwhile, de novo assembly of the metagenomic sequencing data remains challenging for complex communities. To tackle these challenges, we developed a novel algorithm called DRAGoM (Detection of RNA using Assembly Graph from Metagenomic data). DRAGoM first constructs a hybrid graph by merging an assembly string graph and an assembly de Bruijn graph. Then, it classifies paths in the hybrid graph and their constituent readsinto differentncRNA families based on both sequence and structural homology. Our benchmark experiments show that DRAGoMcan improve the performance and robustness over traditional approaches on the classification and quantification of a wide class of ncRNA families.
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Integrated de novo gene prediction and peptide assembly of metagenomic sequencing data
Metagenomics is the study of all genomic content contained in given microbial communities. Metagenomic functional analysis aims to quantify protein families and reconstruct metabolic pathways from the metagenome. It plays a central role in understanding the interaction between the microbial community and its host or environment. De novo functional analysis, which allows the discovery of novel protein families, remains challenging for high-complexity communities. There are currently three main approaches for recovering novel genes or proteins: de novo nucleotide assembly, gene calling and peptide assembly. Unfortunately, their information dependency has been overlooked, and each has been formulated as an independent problem. In this work, we develop a sophisticated workflow called integrated Metagenomic Protein Predictor (iMPP), which leverages the information dependencies for better de novo functional analysis. iMPP contains three novel modules: a hybrid assembly graph generation module, a graph-based gene calling module, and a peptide assembly-based refinement module. iMPP significantly improved the existing gene calling sensitivity on unassembled metagenomic reads, achieving a 92–97% recall rate at a high precision level (>85%). iMPP further allowed for more sensitive and accurate peptide assembly, recovering more reference proteins and delivering more hypothetical protein sequences. The high performance of iMPP can provide a more comprehensive and unbiased view of the microbial communities under investigation. iMPP is freely available from https://github.com/Sirisha-t/iMPP.
more » « less- Award ID(s):
- 1943291
- PAR ID:
- 10401267
- Publisher / Repository:
- Oxford University Press
- Date Published:
- Journal Name:
- NAR Genomics and Bioinformatics
- Volume:
- 5
- Issue:
- 1
- ISSN:
- 2631-9268
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Abstract Motivation De novo transcriptome analysis using RNA-seq offers a promising means to study gene expression in non-model organisms. Yet, the difficulty of transcriptome assembly means that the contigs provided by the assembler often represent a fractured and incomplete view of the transcriptome, complicating downstream analysis. We introduce Grouper, a new method for clustering contigs from de novo assemblies that are likely to belong to the same transcripts and genes; these groups can subsequently be analyzed more robustly. When provided with access to the genome of a related organism, Grouper can transfer annotations to the de novo assembly, further improving the clustering.
Results On de novo assemblies from four different species, we show that Grouper is able to accurately cluster a larger number of contigs than the existing state-of-the-art method. The Grouper pipeline is able to map greater than 10% more reads against the contigs, leading to accurate downstream differential expression analyses. The labeling module, in the presence of a closely related annotated genome, can efficiently transfer annotations to the contigs and use this information to further improve clustering. Overall, Grouper provides a complete and efficient pipeline for processing de novo transcriptomic assemblies.
Availability and implementation The Grouper software is freely available at https://github.com/COMBINE-lab/grouper under the 2-clause BSD license.
Supplementary information Supplementary data are available at Bioinformatics online.
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