RNAhybrid(1) calculate secondary structure hybridisations of RNAs


RNAhybrid [-h] [-b hit_number] [-e energy_cutoff] [-p p-value_cutoff] [-c] [-d xi,theta] [-s set_name] [-f from,to] [-m max_target_length] [-n max_query_length] [-u iloop_upper_limit] [-v bloop_upper_limit] [-g (ps|png|jpg|all)] [-t target_file] [-q query_file] [target] [query]


RNAhybrid is a tool for finding minimum free energy (mfe) hybridisations of a long (target) and a short (query) RNA. The hybridisation is performed in a kind of domain mode, ie. the short sequence is hybridised to the best fitting parts of the long one. The tool is primarily meant as a means for microRNA target prediction. In addition to mfes, the program calculates p-values based on extreme value distributions of length normalised energies.


Give a short summary of command line options.
-b hit_number
Maximal number of hits to show. hit_number hits with increasing minimum free energy (reminder: larger energies are worse) are shown, unless the -e option is used and the energy cut-off has been exceeded (see -e option below) or there are no more hits. Hits may only overlap at dangling bases (5' or 3' unpaired end of target).
Produce compact output. For each target/query pair one line of output is generated. Each line is a colon (:) separated list of the following fields: target name, query name, minimum free energy, position in target, alignment line 1, line 2, line 3, line 4. If a target or a query is given on command line (ie. no -t or -q respectively), its name in the output will be "command line".
-d xi,theta
xi and theta are the position and shape parameters, respectively, of an extreme value distribution (evd). p-values of duplex energies are assumed to be distributed according to such an evd. For a length normalised energy en, we have P[X <= en] = 1 - exp(-exp(-(-en-xi)/theta)), where en = e/log(m*n) with m and n being the lengths of the target and the query, respectively. If the -d option is omitted, xi and theta are estimated from the maximal duplex energy of the query, assuming a linear dependence. The parameters of this linear dependence are coded into the program, but the option -s has to be given to choose from the appropriate set. Note that the evd is mirrored, since good mfes are large negative values.
-s set_name
Used for quick estimate of extreme value distribution parameters (see -d option above). Tells RNAhybrid which target dataset to assume. Valid parameters are 3utr_fly, 3utr_worm and 3utr_human.
-e energy_cutoff
Hits with increasing minimum free energy (reminder: larger energies are worse) less than or equal to energy_cutoff are shown, unless the -b option is used and the number of already reported hits has reached the maximal hit_number (see -b option above). Hits may only overlap at dangling bases (5' or 3' unpaired end of target).
-p p-value_cutoff
Only hits with p-values not larger than p-value_cutoff are reported. See also options -d and -s.
-f from,to
Forces all structures to have a helix from position from to position to with respect to the query. The first base has position 1.
-m max_target_length
The maximum allowed length of a target sequence. The default value is 2000. This option only has an effect if a target file is given with the -t option (see below).
-n max_query_length
The maximum allowed length of a query sequence. The default value is 30. This option only has an effect if a query file is given with the -q option (see below).
-u iloop_upper_limit
The maximally allowed number of unpaired nucleotides in either side of an internal loop.
-v bloop_upper_limit
The maximally allowed number of unpaired nucleotides in a bulge loop.
-g (ps|png|jpg|all)
Produce a plot of the hybridisation, either in ps, png or jpg format, or for all formats together. The plots are saved in files whose names are created from the target and query names ("command_line" if given on the command line). This option only works, if the appropriate graphics libraries are present.
-t target_file
Each of the target sequences in target_file is subject to hybridisation with each of the queries (which either are from the query_file or is the one query given on command line; see -q below). The sequences in the target_file have to be in FASTA format, ie. one line starting with a > and directly followed by a name, then one or more following lines with the sequence itself. Each individual sequence line must not have more than 1000 characters. If no -t is given, either the last argument (if a -q is given) or the second last argument (if no -q is given) to RNAhybrid is taken as a target.
-q query_file
See -t option above.


The energy parameters are taken from:

Mathews DH, Sabina J, Zuker M, Turner DH. "Expanded sequence dependence of thermodynamic parameters improves prediction of RNA secondary structure" J Mol Biol., 288 (5), pp 911-940, 1999

The graphical output uses code from the Vienna RNA package:

Hofacker IL. "Vienna RNA secondary structure server." Nucleic Acids Research, 31 (13), pp 3429-3431, 2003


This man page documents version 2.0 of RNAhybrid.


Marc Rehmsmeier, Peter Steffen, Matthias Hoechsmann.


Character dependent energy values are only defined for [acgtuACGTU]. All other characters lead to values of zero in these cases.


In suboptimal hits, dangling ends appear as Ns if they were in the first or last hybridising position of a previous hit.