FR3D Python

FR3D stands for "Find RNA 3D" and is commonly pronounced "Fred". FR3D enables searching and annotating RNA 3D structures and supports the mmCIF format.

Originally written in Matlab, this repository is a re-implementation of FR3D in Python. FR3D and FR3D Python are developed by the BGSU RNA Bioinformatics group.

Quick start using Docker

Get started with FR3D using containers with Docker, Podman, or Singularity.

Get a Docker image

docker pull ghcr.io/bgsu-rna/fr3d-python
# tag the image with a shorter name for convenience
docker tag ghcr.io/bgsu-rna/fr3d-python fr3d-python

Alternatively, clone the repo and build locally:

git clone https://github.com/BGSU-RNA/fr3d-python
docker build -t fr3d-python .

Notes for M1 Mac users

To build a Docker image on M1 Macs use docker buildx to specify the target platform in order to be able to install all the dependencies:

docker buildx install
docker buildx build -t fr3d-python --platform linux/amd64 .

Annotate RNA 3D structures with basepair interactions
```
docker run -v <path_to_input>:/rna/input -v <path_to_output>:/rna/output fr3d-python python fr3d/classifiers/NA_pairwise_interactions.py -i /rna/input -o /rna/output <pdb_id>
```
where
- path_to_input is the folder where cif files are stored (if a file does not exist already it will downloaded)
- path_to_output is the folder where the results will be generated
- pdb_id is the PDB identifier of the structure to be annotated (for example, 1S72). Multiple PDB identifiers can be specified
- Optional flag determines which output files are generated
  - -c basepair outputs a file listing basepairs in the 12 Leontis-Westhof families
  - -c basepair_detail outputs Leontis-Westhof pairs with more detail about asymmetric pairs and with bifurcated and alternative pairs
  - -c sO outputs a file with base-oxygen stacking annotations
  - -c basepair,cO outputs both basepair and sO annotation files; don't put spaces between the category types

Notes on updating Docker image in Container registry

The Docker images are hosted on the GitHub Container registry. Currently the images need to be updated manually as follows:

Authenticate using Personal Access Token (required only once)

Create and push a new image

docker build -t ghcr.io/bgsu-rna/fr3d-python .
docker push ghcr.io/bgsu-rna/fr3d-python

In the future this should be done automatically with GitHub Actions.

Understanding the output

Note that as of April 2022, NA_pairwise_interactions.py outputs annotations of RNA basepairs, including basepairs made by some modified nucleotides, but this is in a test phase, and not all basepairs are being annotated. Work to refine the basepair annotations will continue in Summer 2022.

The program NA_pairwise_interactions.py generates tab-separated files called <pdb_id>_basepairs.txt that list basepair interactions between pairs of nucleotides (an RNA 3D structure can contain 0 or more basepairs). The files contain the following fields:

Unit ID of the first nucleotide in the Unit ID format
basepair interaction between the first and second nucleotides, defined according to the Leontis-Westhof nomenclature
Unit ID of the second nucleotide
number of interactions crossed by the basepair (0 for nested basepairs or an integer for non-nested basepairs)

For example:

1S72|1|0|A|1193	tSH	1S72|1|0|U|1205	0
1S72|1|0|A|631	tSH	1S72|1|0|U|2607	25

Installation without Docker

Clone FR3D Python

git clone https://github.com/BGSU-RNA/fr3d-python
cd fr3d-python
python setup.py install

⚠️ Any time files such as fr3d/cif/reader.py get edited, you need to rerun the python setup.py install command.

Set up on Python 2.7

fr3d-python requires PDBx. Follow instructions at wwPDB to install PDBx on your local machine and place in your Python path.

Set up on Python 3.x

In order to use FR3D in Python 3, you will need to install the mmcif-pdbx package:

pip install mmcif-pdbx

This will put the PDBx package in your Python path.

Solving TypeError when reading .cif files

On Windows, mmcif-pdbx can experience a TypeError when loading .cif files. This is apparently because your machine expects the iterator loop to deal with bytes. Try this: find where mmcif-pdbx is installed using pip show mmcif-pdbx. Edit C:[python-path]\Lib\site-packages\pdbx\reader.py At lines 387 and 397, after for line in file_iterator: add a line line = line.decode().

Note for Windows users about case-sensitive filenames

FR3D Python uses case-sensitive file names because chain identifiers in PDB are case sensitive. Windows looks like it uses case sensitive filenames, but if you create a file called Data.txt and then save another file in the same place called data.txt, it will overwrite Data.txt and it will be listed as Data.txt because that was there first. Recent versions of Windows allow for case sensitivity.

For FR3D Python, you need to enable case sensitivity in filenames in the units folder. See https://docs.microsoft.com/en-us/windows/wsl/case-sensitivity to read about how to enable case sensitivity for a specific directory.

The key steps seem to be: - Enable-WindowsOptionalFeature -Online -FeatureName Microsoft-Windows-Subsystem-Linux - fsutil.exe file setCaseSensitiveInfo <path to folder> enable

Note that the first step is not mentioned in the Microsoft documentation.

Running FR3D Python

Navigate to the classifiers folder and run the following command to ensure that the installation was successful:

python NA_pairwise_interactions.py --help

You can update localpath.py to represent your file structure. Alternatively, use the --input (-i) and --output (-o) flags to specify the location of your input cif files and the output text files.

For example, if you already have the .cif file downloaded, use the --input flag to specify the directory your cif file is in:

python NA_pairwise_interactions.py --input <path_to_input> <pdb_id>

Additionally, if you don't have the cif file in your localpath, you can run

python NA_pairwise_interactions.py <pdb_id>.cif

and this will download the file for processing and run the program.

Output can be specified using the -o or --output tags, for example:

python NA_pairwise_interactions.py --output <path_to_output> <pdb_id>

This will write where you wish to output your annotations. If the specified directory doesn't exist, one will be made.

Get in touch

If you have any questions or feature requests, feel free to submit an issue or get in touch via a contact form on the website of the BGSU RNA Bioinformatics group.

mvaradi/fr3d-python