FastOMA

FastOMA is a scalable software package for inferring orthology relationship.

Input and Output:

Input:

1- Sets of protein sequences in FASTA format (with .fa extension) in the folder proteome. The name of each fasta file is the name of species. Please make sure that the name of fasta records do not contain ||.

2- The omamer database which you can download this which is from OMA browser. This file is 13 Gb containing all the gene families of the Tree of Life or you can download it for a subset of them, e.g. Primates (352MB).

3- Sepecies tree in newick format. Note that the name of leaves of the tree (species name) should be the same as the file name of fastas (without .fa extension) (item 1). You can see an example in the testdata folder.

$ ls proteome
AQUAE.fa  CHLTR.fa  MYCGE.fa
$ cat species_tree.nwk
((AQUAE,CHLTR)inter1,MYCGE)inter2;

Besides, the internal node should not contain any special character (e.g. \ / or space). The reason is that FastOMA write some files whose names contains the internal node's name. If the species tree does not have label for some/all internal nodes, FastOMA labels them sequentially.

Output:

Orthology information as HOG strcutre in OrthoXML format which can be used with PyHAM

How to run FastOMA

In summary, you need to 1) install FastOMA using pip after its prerequisites (below), and 2) put the input files in the folder in_folder and 3) run FastOMA using the nextflow recipe FastOMA_script.nf.

python -m pip install -e ./FastOMA 
nextflow  FastOMA_script.nf  --input_folder /path/to/in_folder   --output_folder /path/to/out_folder 

For a detailed instruction start from prerequisites and continue to the section How to run FastOMA on the test data (details).

prerequisites

FastOMA needs following software packages: omamer, Nextflow, Biopython, dendropy, pyparsing , ete3, fasttree and mafft.

To do so, you can start with a fresh conda environment.

conda create --name FastOMA python=3.9
conda activate FastOMA

You can install the packages using pip. You can always make sure whether you are using the python that you intended to use with which python and which python3.

python -m pip install --upgrade pip
python -m pip install biopython
python -m pip install omamer
# python -m pip install pytables==3.6.1 # if you had trouble with pytables for omamer.
python -m pip install ete3  
python -m pip install nextflow
python -m pip install pyparsing
python -m pip install DendroPy
python -m pip install future
python -m pip install lxml

You may use conda to install mafft, fasttree and openjdk (the alternative for Java 11< version <17 which is needed for nextflow).

conda install -c bioconda mafft fasttree
conda install -c conda-forge openjdk

You may also need to have the following for nextflow to work.

JAVA_HOME="/path/to/jdk-17"
NXF_JAVA_HOME="/path/to/jdk-17"
export PATH="/path/to/jdk-17/bin:$PATH"

You can make sure that omamer and nextflow is installed with running

omamer
nextflow -h

How to run FastOMA on the test data (details)

First, download the FastOMA package:

wget https://github.com/DessimozLab/FastOMA/archive/refs/heads/master.zip
unzip master.zip
mv FastOMA-master FastOMA

or clone it

git clone [email protected]:DessimozLab/FastOMA.git

Then install it

ls FastOMA/setup.py
python -m pip install -e FastOMA 

The output would be

  Preparing metadata (setup.py) ... done
Building wheels for collected packages: FastOMA
  Building wheel for FastOMA (setup.py) ... done
  Created wheel for FastOMA: filename=FastOMA-0.0.5-py3-none-any.whl size=29386 sh
Successfully built FastOMA
Installing collected packages: FastOMA
Successfully installed FastOMA-0.0.5

You can check your installation with

infer-roothogs --version

Then, cd to the testdata folder and download the omamer database and change its name to omamerdb.h5.

cd FastOMA/testdata
wget https://omabrowser.org/All/Primates.h5    # 352MB
mv Primates.h5  in_folder/omamerdb.h5 

(This is for the test however, I would suggest downloading the LUCA.h5 instead of Primates.h5 for your real analysis.). Check the item 2 in the input section for details.

Now we have such a structre in our testdata folder.

$ tree ../testdata/
├── in_folder
│   ├── omamerdb.h5
│   ├── proteome
│   │   ├── AQUAE.fa
│   │   ├── CHLTR.fa
│   │   └── MYCGE.fa
│   └── species_tree.nwk
└── README.md

Finally, run the package using nextflow as below:

# cd FastOMA/testdata
nextflow ../FastOMA_script.nf  --input_folder in_folder   --output_folder out_folder  -with-report

Note that to have a comprehensive test, we set the default value of needed cpus as 10.

After few minutes, the run for test data finishes.

[] process > omamer_run (3)      [100%] 3 of 3 ✔
[] process > infer_roothogs (1)  [100%] 1 of 1 ✔
[] process > batch_roothogs (1)  [100%] 1 of 1 ✔
[] process > hog_big (1)         [100%] 1 of 1 ✔
[] process > hog_rest ( )        [100%] 2 of 2 ✔
[] process > collect_subhogs (1) [100%] 1 of 1 ✔

If the run interrupted, by adding -resume to the nextflow commond line, you can continue your previous nextflow job.

Then, following files and folders should appear in the folder out_folder which was the argument.

$ tree -L 1  out_folder
├── hogmap
├── output_hog.orthoxml
├── pickle_rhogs
├── rhogs_all
├── rhogs_big
└── rhogs_rest

among which output_hog_.orthoxml is the final output. Its content looks like this

<?xml version="1.0" ?>
<orthoXML xmlns="http://orthoXML.org/2011/" origin="OMA" originVersion="Nov 2021" version="0.3">
   <species name="MYCGE" NCBITaxId="1">
      <database name="QFO database " version="2020">
         <genes>
            <gene id="1000000000" protId="sp|P47500|RF1_MYCGE"/>
            <gene id="1000000001" protId="sp|P13927|EFTU_MYCGE"/>
            <gene id="1000000002" protId="sp|P47639|ATPB_MYCGE"/>
            
 ...
      <orthologGroup id="HOG:B0885011_sub10003">
         <property name="TaxRange" value="inter1"/>
         <geneRef id="1002000004"/>
         <geneRef id="1001000004"/>
      </orthologGroup>
   </groups>
</orthoXML>

using omamer's output

The first step of the FastOMA pipele is to run OmaMer. If you already have the hogmap files, you can put them in the in_folder/hogmap_input_folder. Then your structure of files will be

$ tree ../testdata/
├── in_folder
│   ├── hogmap_input_folder
│   │   ├── CHLTR.fa.hogmap
│   │   ├── MYCGE.fa.hogmap
│   ├── omamerdb.h5
│   ├── proteome
│   │   ├── AQUAE.fa
│   │   ├── CHLTR.fa
│   │   └── MYCGE.fa
│   └── species_tree.nwk
└── README.md

In this case, FastOMA uses two hogmap files and only run omamer for the AQUAE.fa. Then continue the rest of pipeline. Let's save the planet together with green computational Biology.

Run on a cluster

For running on a SLURM cluster you can add -c ../nextflow_slurm.config to the commond line.

# cd FastOMA/testdata
# rm -r out_folder work          # You may remove stuff from previous run
# ls ../FastOMA_script.nf 

nextflow ../FastOMA_script.nf  -c ../nextflow_slurm.config   --input_folder in_folder   --output_folder out_folder

You may need to re-run nextflow command line by adding -resume, if the allocated time is not enough for your dataset.

You may need to increase the number of opoened files in your system with ulimit -n 131072 or higher.

If you have the omamer mappings, you can put them in hogmap folder and run the following

$ ls
hogmap  in_folder  genetrees
$ nextflow   FastOMA/archive/FastOMA_script_after_omamer.nf   --input_folder in_folder   --output_folder out_folder

for i in $(ls .); do mv $i `basename $i`.fa.hogmap; done 

Change log

• prelease
• prelease v.0.0.6: use --fragment-detection for infer-subhogs and --low-so-detection --fragment-detection
• prelease v.0.0.6: using input hogmpa
• prelease v.0.0.5: adding pip setup.py
• prelease v.0.0.4: simple nextflow
• prelease v.0.0.3: with dask