Rosetta 3.4
Documentation for RNA loop modeling (lock-and-key problem) with Stepwise Assembly, using swa_rna_main and swa_rna_util executables
Parin Sripakdeevong, Rhiju Das


March 2012 by Parin Sripakdeevong (sripakpa [at] and Rhiju Das (rhiju [at]

Code and Demos

This code builds single-stranded RNA loops using a deterministic, enumerative sampling method called Stepwise Assembly. A demo example for building the first (5' most) nucleotide of a 6-nucleotide loop is given in rosetta_demos/public/SWA_RNA_Loop/. The same protocol can then be recursively applied to build the remaining nucleotides in the loop, one individual nucleotide at a time.

(Note: the Stepwise Assembly method constructs full-length RNA loops through the recursive building of each individual RNA nucleotides over multiple steps. The enumerative nature of the method makes the full-calculation computationally expensive, requiring for example 15,000 CPU hours to build a single 6-nucleotide RNA loop. While this full-calculation is now feasible on a high-performance computer clusters, perform the full-calculation in the demo would be too excessive.)

The central codes are located in the src/protocols/swa_rna/ folder. The applications are in apps/public/swa_rna_main and apps/public/swa_rna_util


Sripakdeevong, P., Kladwang, W. & Das, R. (2012), “An enumerative stepwise ansatz enables atomic-accuracy RNA loop modeling”, Proc Natl Acad Sci USA. doi:10.1073/pnas.1106516108


This method builds single-stranded RNA loops using a deterministic, enumerative sampling method called Stepwise Assembly. The modeling situation considered here is the lock-and-key problem. Given a template PDB that contains nucleotides surrounding a missing RNA loop, the Stepwise Assembly method finds the loop conformation (the key) that best fits the surrounding structure (the lock).



There is only one mode to run SWA_RNA_Loop at present.

Input Files

Required file

You need two files:

additional files:

Preprocessing of input files

Input PDB file can be converted into the Rosetta RNA PDB format using the following command:

	rosetta_tools/SWA_RNA_python/SWA_dagman_python/misc/ IN_PDB.pdb -output_pdb OUT_PDB.pdb

Replace 'IN_PDB.pdb' with your input PDB filename. Replace 'OUT_PDB.pdb' with the filename you want the converted PDB to be outputted to.

How to run the job

The SWA_RNA_python package located at rosetta_tools/SWA_RNA_python/ contains the scripts necessary to setup and run the Stepwise Assembly protocol. Instructions are provided in steps 1)-4) below:

1) Specify the location of the rosetta bin folder and rosetta database folder by editing the file rosetta_tools/SWA_RNA_python/SWA_dagman_python/utility/

For example, if the main rosetta folder is located at ~/rosetta/, then the file should look as follow:



2) Add the SWA_RNA_python package location to the PYTHON path. For bash shell users, the location can be directly added to the ~/.bashrc file:

	export PYTHONPATH=$PYTHONPATH:~/rosetta/rosetta_tools/SWA_RNA_python/

3) After the paths are correctly specified, the following command is used to setup everything needed run the Stepwise Assembly job:

	rosetta_tools/SWA_RNA_python/SWA_dagman_python/SWA_DAG/ -s template.pdb -fasta fasta -sample_res 3-8 -single_stranded_loop_mode True -local_demo True -native_pdb native.pdb
	The "-s" flag specifies the @ref template_PDB file
	The "-fasta" flag specifies the @ref fasta file
	The "-sample_res" flag specifies the sequence number of nucleotides in the missing loop. For example, 3-8 means that the missing loop nucleotides are located at sequence position 3 4 5 6 7 and 8.
	The "-single_stranded_loop_mode" flag specifies that the job involve modeling a single-stranded loop (i.e. the lock-and-key problem).
	The "-local_demo" flag indicate that this is demo to be run on a local laptop or desktop. The calculation perform here is to only build the first (5' most) nucleotide of the 6-nucleotides RNA loop.
	The "-native_pdb" flag specifies the @ref native_PDB file and is optional. 

4) Type "source LOCAL_DEMO" to execute the Rosetta protocol.

The provided instruction will allow the user to build the first (5' most) nucleotide of a N-nucleotide loop. As previously stated, the full-calculation to build full-length RNA loops is quite computationally expensive and is beyond the scope of this documentation. The SWA_RNA_python package is, however, equipped to run this recursive full-calculation on a high-performance computer clusters. The package utilize concept familiar from the Map/Reduce Direct Acyclic Graph framework to order the calculation steps and allocate resources to recursive build the full-length RNA loop over multiple steps, one individual RNA nucleotide at a time. If any user is interested, please contact Parin Sripakdeevong (sripakpa [at] and we will be happy to provide additional instructions.

Expected Outputs

The expected outputs are two silent_files:

	A) region_0_1_sample.out: This silent_file contain 108 structures, corresponding to the 108 lowest energy conformations.
	B) region_0_1_sample.cluster.out: Same as A) but after clustering of the models to remove redundant conformations.

Post Processing

In both silent_files, the total energy score is found under the 'score' column. If the "native_pdb" flag was included, then the RMSD (in angstrom units) between the native_pdb and each Rosetta model is found under the 'NAT_rmsd' column.

Finally, use the following command to extract the top 5 energy cluster centers:

	rosetta_tools/SWA_RNA_python/SWA_dagman_python/misc/ -tag S_0 S_1 S_2 S_3 S_4  -silent_file region_0_1_sample.cluster.out

After running the command, the extracted PDB files should appear in the pose_region_0_1_sample.cluster.out/ subfolder.

New things since last release

This application is new as of Rosetta 3.4.

 All Classes Namespaces Files Functions Variables Typedefs Enumerations Enumerator Friends Defines