MutantHuntWGS: A Pipeline for Identifying Saccharomyces cerevisiae Mutations

MutantHuntWGS is a user-friendly pipeline for analyzing Saccharomyces cerevisiae whole-genome sequencing data. It uses available open-source programs to: (1) perform sequence alignments for paired and single-end reads, (2) call variants, and (3) predict variant effect and severity. MutantHuntWGS outputs a shortlist of variants while also enabling access to all intermediate files. To demonstrate its utility, we use MutantHuntWGS to assess multiple published datasets; in all cases, it detects the same causal variants reported in the literature. To encourage broad adoption and promote reproducibility, we distribute a containerized version of the MutantHuntWGS pipeline that allows users to install and analyze data with only two commands. The MutantHuntWGS software and documentation can be downloaded free of charge from https://github.com/mae92/MutantHuntWGS.

[1]  Matthew W Vaughn,et al.  Mapping epigenetic mutations in fission yeast using whole-genome next-generation sequencing. , 2009, Genome research.

[2]  Helga Thorvaldsdóttir,et al.  Integrative Genomics Viewer (IGV): high-performance genomics data visualization and exploration , 2012, Briefings Bioinform..

[3]  Pablo Cingolani,et al.  © 2012 Landes Bioscience. Do not distribute. , 2022 .

[4]  Rommie E. Amaro,et al.  Comparative chemical genomics reveal that the spiroindolone antimalarial KAE609 (Cipargamin) is a P-type ATPase inhibitor , 2016, Scientific Reports.

[5]  W. Krzyzosiak,et al.  Sequence-non-specific effects of RNA interference triggers and microRNA regulators , 2009, Nucleic acids research.

[6]  Rommie E. Amaro,et al.  Rapid Chagas Disease Drug Target Discovery Using Directed Evolution in Drug-Sensitive Yeast. , 2017, ACS chemical biology.

[7]  Fred Winston,et al.  Whole‐Genome Sequencing of Yeast Cells , 2019, Current protocols in molecular biology.

[8]  Jamal Tazi,et al.  Regulated functional alternative splicing in Drosophila , 2011, Nucleic acids research.

[9]  Michael F. Wangler,et al.  Model Organisms Facilitate Rare Disease Diagnosis and Therapeutic Research , 2017, Genetics.

[10]  Fred Winston,et al.  Back to the Future: Mutant Hunts Are Still the Way To Go , 2016, Genetics.

[11]  Heeseung Jo,et al.  Faster single-end alignment generation utilizing multi-thread for BWA. , 2015, Bio-medical materials and engineering.

[12]  Corey Nislow,et al.  Complementation of Yeast Genes with Human Genes as an Experimental Platform for Functional Testing of Human Genetic Variants , 2015, Genetics.

[13]  A. Hamza,et al.  Cross-Species Complementation of Nonessential Yeast Genes Establishes Platforms for Testing Inhibitors of Human Proteins , 2020, Genetics.

[14]  Alfred V. Aho,et al.  Awk — a pattern scanning and processing language , 1979, Softw. Pract. Exp..

[15]  Carl Boettiger,et al.  An introduction to Docker for reproducible research , 2014, OPSR.

[16]  Daniel Blankenberg,et al.  CloudMap: A Cloud-Based Pipeline for Analysis of Mutant Genome Sequences , 2012, Genetics.

[17]  E. Mardis Next-generation sequencing platforms. , 2013, Annual review of analytical chemistry.

[18]  Sven Rahmann,et al.  Snakemake--a scalable bioinformatics workflow engine. , 2012, Bioinformatics.

[19]  Gonçalo R. Abecasis,et al.  The variant call format and VCFtools , 2011, Bioinform..

[20]  Steven L Salzberg,et al.  Fast gapped-read alignment with Bowtie 2 , 2012, Nature Methods.

[21]  Fred Winston,et al.  A conserved genetic interaction between Spt6 and Set2 regulates H3K36 methylation , 2018, bioRxiv.

[22]  Thomas E Wilson,et al.  Discovery of Mutations in Saccharomyces cerevisiae by Pooled Linkage Analysis and Whole-Genome Sequencing , 2010, Genetics.

[23]  Daniel J. Blankenberg,et al.  Galaxy: A Web‐Based Genome Analysis Tool for Experimentalists , 2010, Current protocols in molecular biology.

[24]  Gabor T. Marth,et al.  Rapid whole-genome mutational profiling using next-generation sequencing technologies. , 2008, Genome research.

[25]  V. Lundblad,et al.  Cloning Yeast Genes by Complementation , 1989, Current protocols in molecular biology.

[26]  Austin G. Meyer,et al.  Systematic humanization of yeast genes reveals conserved functions and genetic modularity , 2015, Science.

[27]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[28]  Daniel J. Blankenberg,et al.  Galaxy: a platform for interactive large-scale genome analysis. , 2005, Genome research.

[29]  Edith D. Wong,et al.  The Reference Genome Sequence of Saccharomyces cerevisiae: Then and Now , 2013, G3: Genes, Genomes, Genetics.

[30]  Hanlee P. Ji,et al.  Next-generation DNA sequencing , 2008, Nature Biotechnology.

[31]  M. DePristo,et al.  The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. , 2010, Genome research.

[32]  Edith D. Wong,et al.  Saccharomyces Genome Database: the genomics resource of budding yeast , 2011, Nucleic Acids Res..

[33]  Heng Li,et al.  A statistical framework for SNP calling, mutation discovery, association mapping and population genetical parameter estimation from sequencing data , 2011, Bioinform..

[34]  Pablo Prieto,et al.  The impact of Docker containers on the performance of genomic pipelines , 2015, PeerJ.

[35]  P. Ng,et al.  SIFT missense predictions for genomes , 2015, Nature Protocols.

[36]  Yasukazu Nakamura,et al.  Mudi, a web tool for identifying mutations by bioinformatics analysis of whole‐genome sequence , 2014, Genes to cells : devoted to molecular & cellular mechanisms.

[37]  D. Botstein,et al.  Yeast: An Experimental Organism for 21st Century Biology , 2011, Genetics.

[38]  David Botstein,et al.  Analysis of Polygenic Mutants Suggests a Role for Mediator in Regulating Transcriptional Activation Distance in Saccharomyces cerevisiae , 2015, Genetics.

[39]  H. Hakonarson,et al.  ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data , 2010, Nucleic acids research.

[40]  Jason N Berman,et al.  How Surrogate and Chemical Genetics in Model Organisms Can Suggest Therapies for Human Genetic Diseases , 2018, Genetics.