An optimal and non-overlapping DNA repeat searcher.
Includes tools for dealing poly-purine/poly-pyrimidine tracts (PPTs).
What does that mean?
What it means is that
agate will search of repeats in DNA sequences, but
unlike most repeat finder tools, it will only return the best matches and any
sequence segment can only be associated with one reported repeat.
There is a bunch of extra work in trying to select the optimal sequence and allow for mismatch errors. Most repeat finders show you a whole bunch of candidate matches, and sometimes you just want the best. The selection of these can in turn impact the selection of neighbouring repeat sequences etc.
For the particular research problem that the software was created to address, no overlaps was a condition.
This repo contains source for four executables. They are probably not master pieces of software engineering as I wrote them in my free time while doing a PhD in an unrelated area, and the requirements changes substantially while developing each tool:
- repeat-finder: Main program for finding optimal and exclusive repeating motifs using mismatches. Further description found below.
- ppt-se: Looks for poly-purine/poly-pyrimidine tracts (PPTs) with an internal axis of symmetry. This includes, for example, AAAGAAA, GAGGAG. It has the additional commands "sel" and "seu" which specify the size limits of the symmetrical element.
- ppt-mismatch: Has the additional capability of finding PPT arrays with mismatches, the "e" command specified the block size within which only one mismatch is allowed. Mismatches are also disallowed within the terminal two nucleotides.
- ppt: Looks for PPTs eg AAAAAAAA, AGGGGA, GGGGG, CCCCCC, CCTCTTTCC, TTTTTT. "l" and "u" are size limiters for the overall tract size.
- expected: Calculates the expected number of repeats based on the equations in the paper: de Wachter, Rupert (1981). The Number of Repeats Expected in Random Nucleic Acid Sequences and Found in Genes. J. theor. Biol. 91, 71-98
RepeatFinder has been developed in C with a minimal of dependencies. It should be compilable on any operating environment that supports the standard ANSI C library.
By default the compilation system is setup to support the Make build system supported by the GCC compiler
Simply typing make in the source directory should compile three executables:
agate-repeat-finder, ppt, ppt-mismatch, ppt-se, expected
If you don't have the Make build system you will have to compile and link the source code yourself. You may need to check the Makefile environment variables to see that they conform to your setup.
Instructions for use
Create a database of matches.
This is done with the "-d" option followed by a number indicating the maximum size of motifs to look for. e.g.
$ repeat-finder.exe -d 3 chr01.fsa
This will look for motifs of size 3 and below in the file "chr01.fsa".
This is the most time consuming part and once you have run it you shouldn't have to run it again unless you want to change the maximum size of motifs or you get a new version of the program. The database filename is the same as the sequence with ".database" appended. So for the above example, a database file called "chr01.fsa.database" is created.
The database file must be the same as the sequence filename with ".database" appended. So if you rename the sequence file, you must rename the database file or regenerate it.
Changing the error distance.
By default databases are created with a minimum error distance of 5. If you wish to alter this you can also specify an error distance when you create a database using the "-e" option. e.g.
$ repeat-finder.exe -d 3 -e 7 chr01.fsa
Sets the minimum error distance to be 7 nucleotides.
Specify what matches you are interested in using the following:
-f <size> - Output the <size> flanking bases to a match (default 50) -n <number> - search for motifs of size <number>. You can use this option more than once, for example to specify motifs of sizes 1 and 2 you can use "-n 2 -n 1" -m <motif> - search for a particular motif. e.g "-m ATT". You can also use this more than once. -l <lower> - find matches with at least <lower> repeats. -u <upper> - ignore matches with more than <upper> repeats. -lp <lower> - lowest purity repeats to find -up <upper> - greatest purity repeats to find (Note: range of purities is constrained by error distance)
Results are output to the file "results.csv". Be warned that this is overwritten if it already exists so rename results you wish to keep.
Use "-cr" to look for compound repeats, and "-gap" to specify the maximum gap between repeats. If you only are interested in degenerate repeats then use "-dr" as "-cr" looks for both compound and degenerate repeats.
You also must use the options for normal matches to specify which ones to output. e.g.
$ repeat-finder -cr -gap 4 -n 2 chr01.fsa
Looks for compound repeats with a gap of at most 4. It then filters these for only those that have at least one sub-repeat with a motif of size 2.
Results are output to the file "results.csv". The compound matches are reported at the end, and also has a column to indicate the the indexes of the sub repeats.
Extracting to FASTA
To output to a fasta file use "-i" option along with the index of the match. If the index is of a compound repeat you must also specify either -cr or -dr (depending on what you used to get the index) and the same -gap number on the command line.
You can use the index "-1" to indicate that all repeats should be output to fasta.
The file created is by default called "repeats.fsa"
What research has this been used in?
- Bagshaw, A., Pitt, J.P.W., Gemmell, N.J. (2008). High frequency of microsatellites in S. cerevisiae meiotic recombination hotspots. BMC Genomics 9:49 doi:10.1186/1471-2164-9-49
- Bagshaw, A., Pitt, J.P.W., Gemmell, N.J. (2006). Association of poly-purine/poly-pyrimidine sequences with meiotic recombination hot spots. BMC Genomics 7:179, doi:10.1186/1471-2164-7-179
And I've heard it's been used by a few other people in the same lab, although perhaps not to a sufficient extent to be referenced.
Code repository on github: https://github.com/ferrouswheel/agate
NOTE: This project used to be called repeatfinder and be hosted on sourceforge, but then they broke the drupal website and I'm more fond of github. The reason for the name change is that there are many other pieces of software called "repeatfinder" that do the same thing. "Agate" is a satellite/missle name and was the shortest name mostly composed of nucleotide letters ;-)