# Evolutionary Analysis

# Live Resources

usegalaxy.org usegalaxy.eu usegalaxy.org.au usegalaxy.be usegalaxy.fr
workflow workflow run run workflow workflow run run workflow workflow run run workflow workflow run run workflow workflow run run
history history view view history history view view history history view view history history view view history history view view

# What's the point?

Wu et al. showed recombination between COVID-19 and bat coronaviruses located within the S-gene. We want to confirm this observation and provide a publicly accessible workflow for recombination detection.

In previous coronavirus outbreaks (SARS), retrospective analyses determined that adaptive substitutions might have occurred in the S-protein Zhang et al., e.g., related to ACE2 receptor utilization. While data on COVID-19 are currently limited, we investigated whether or not the lineage leading to them showed any evidence of positive diversifying selection.

# Outline

We employ a recombination detection algorithm (GARD) developed by Kosakovsky Pond et al. and implemented in the hyphy package. To select a representative set of S-genes we perform a blast search using the S-gene CDS from NC_045512 as a query against the nr database. We select coding regions corresponding to the S-gene from a number of COVID-19 genomes and original SARS isolates. This set of sequences can be found in this repository

We then generate a codon-based alignment using the workflow shown below and perform the recombination analysis using the gard tool from the hyphy package.

For selection analyses, we apply the Adaptive Branch Site Random Effects method to test whether or each branch of the tree shows evidence of diversifying positive selection along a fraction of sites using the absrel tool from the hyphy package.

# Inputs

A set of unaligned CDS sequences for the S-gene.

# Outputs

A recombination report:

and a map of possible recombination hotspots:

A selection analysis summary and tree (COVID-19 isolate is MN988668_1)

and a plot of the inferred ω distribution for the MN988668_1 branch.

# History and workflow

A Galaxy workspace (history) containing the most current analysis can be imported from here.

The publicly accessible workflow can be downloaded and installed on any Galaxy instance. It contains version information for all tools used in this analysis.

The workflow takes unaligned CDS sequences, translates them with EMBOSS:tanseq, aligns translations using mafft, realigns original CDS input using the mafft alignment as a guide and sends this codon-based alignment to gard.

# BioConda

Tools used in this analysis are also available from BioConda:

Name Link
emboss Anaconda-Server Badge
mafft Anaconda-Server Badge
hyphy Anaconda-Server Badge
fasttree Anaconda-Server Badge