3 .\" DO NOT MODIFY THIS FILE! It was generated by help2man 1.38.2.
4 .TH RNAALIFOLD "1" "July 2013" "RNAalifold 2.1.2" "User Commands"
6 RNAalifold \- manual page for RNAalifold 2.1.2
9 [\fIoptions\fR] \fI<file1.aln>\fR
13 calculate secondary structures for a set of aligned RNAs
15 Read aligned RNA sequences from stdin or file.aln and calculate their minimum
16 free energy (mfe) structure, partition function (pf) and base pairing
17 probability matrix. Currently, the input alignment has to be in CLUSTAL format.
18 It returns the mfe structure in bracket notation, its energy, the free energy
19 of the thermodynamic ensemble and the frequency of the mfe structure in the
20 ensemble to stdout. It also produces Postscript files with plots of the
21 resulting secondary structure graph ("alirna.ps") and a "dot plot" of the
22 base pairing matrix ("alidot.ps"). The file "alifold.out" will contain a
23 list of likely pairs sorted by credibility, suitable for viewing with
24 "AliDot.pl". Be warned that output file will overwrite any existing files of
27 \fB\-h\fR, \fB\-\-help\fR
30 \fB\-\-detailed\-help\fR
31 Print help, including all details and hidden
35 Print help, including hidden options, and exit
37 \fB\-V\fR, \fB\-\-version\fR
38 Print version and exit
39 .SS "General Options:"
41 Below are command line options which alter the general behavior of this
44 \fB\-C\fR, \fB\-\-constraint\fR
45 Calculate structures subject to constraints.
46 The constraining structure will be read from
47 \&'stdin', the alignment has to be given as a
48 file name on the command line.
52 The program reads first the sequence, then a string containing constraints on
53 the structure encoded with the symbols:
55 \&. (no constraint for this base)
57 | (the corresponding base has to be paired
59 x (the base is unpaired)
61 < (base i is paired with a base j>i)
63 \f(CW> (base i is paired with a base j<i)\fR
65 and matching brackets ( ) (base i pairs base j)
67 With the exception of "|", constraints will disallow all pairs conflicting
68 with the constraint. This is usually sufficient to enforce the constraint,
69 but occasionally a base may stay unpaired in spite of constraints. PF folding
70 ignores constraints of type "|".
73 Produce a colored version of the consensus
74 strcture plot "alirna.ps" (default b&w
80 Produce a colored and structure annotated
81 alignment in PostScript format in the file
82 "aln.ps" in the current directory.
87 Do not produce postscript output
92 Select additional algorithms which should be included in the calculations.
93 The Minimum free energy (MFE) and a structure representative are calculated
96 \fB\-p\fR, \fB\-\-partfunc\fR[=\fIINT\fR]
97 Calculate the partition function and base
98 pairing probability matrix in addition to the
99 mfe structure. Default is calculation of mfe
104 In addition to the MFE structure we print a coarse representation of the pair
105 probabilities in form of a pseudo bracket notation, followed by the ensemble
106 free energy, as well as the centroid structure derived from the pair
107 probabilities together with its free energy and distance to the ensemble.
108 Finally it prints the frequency of the mfe structure.
110 An additionally passed value to this option changes the behavior of partition
111 function calculation:
112 \fB\-p0\fR deactivates the calculation of the pair probabilities, saving about 50%
113 in runtime. This prints the ensemble free energy \fB\-kT\fR ln(Z).
115 \fB\-\-MEA\fR[=\fIgamma\fR]
116 Calculate an MEA (maximum expected accuracy)
121 If gamma is not specified a default of gamma=1 is used.
122 Using \fB\-\-MEA\fR implies \fB\-p\fR
123 See also RNAfold man page for details.
126 Output "most informative sequence" instead of
127 simple consensus: For each column of the
128 alignment output the set of nucleotides with
129 frequence greater than average in IUPAC
134 \fB\-s\fR, \fB\-\-stochBT\fR=\fIINT\fR
135 Stochastic backtrack. Compute a certain number
136 of random structures with a probability
137 dependend on the partition function. See \fB\-p\fR
140 \fB\-\-stochBT_en\fR=\fIINT\fR
141 same as "\-s" but also print out the energies
142 and probabilities of the backtraced
145 \fB\-S\fR, \fB\-\-pfScale\fR=\fIscaling\fR factor
146 In the calculation of the pf use scale*mfe as
147 an estimate for the ensemble free energy
148 (used to avoid overflows).
150 The default is 1.07, useful values are 1.0 to 1.2. Occasionally needed for
152 You can also recompile the program to use double precision (see the README
155 \fB\-c\fR, \fB\-\-circ\fR
156 Assume a circular (instead of linear) RNA
161 \fB\-\-bppmThreshold=\fR<value>
162 Set the threshold for base pair probabilities
163 included in the postscript output
167 By setting the threshold the base pair probabilities that are included in the
168 output can be varied. By default only those exceeding 1e\-5 in probability
169 will be shown as squares in the dot plot. Changing the threshold to any other
170 value allows for increase or decrease of data.
172 \fB\-g\fR, \fB\-\-gquad\fR
173 Incoorporate G\-Quadruplex formation into the
174 structure prediction algorithm
179 \fB\-T\fR, \fB\-\-temp\fR=\fIDOUBLE\fR
180 Rescale energy parameters to a temperature of
181 temp C. Default is 37C.
183 \fB\-4\fR, \fB\-\-noTetra\fR
184 Do not include special tabulated stabilizing
185 energies for tri\-, tetra\- and hexaloop
186 hairpins. Mostly for testing.
190 \fB\-d\fR, \fB\-\-dangles\fR=\fIINT\fR
191 How to treat "dangling end" energies for
192 bases adjacent to helices in free ends and
197 With \fB\-d2\fR dangling energies will be added for the bases adjacent to a helix on
202 The option \fB\-d0\fR ignores dangling ends altogether (mostly for debugging).
205 Produce structures without lonely pairs
206 (helices of length 1).
210 For partition function folding this only disallows pairs that can only occur
211 isolated. Other pairs may still occasionally occur as helices of length 1.
214 Do not allow GU pairs
218 \fB\-\-noClosingGU\fR
219 Do not allow GU pairs at the end of helices
223 \fB\-\-cfactor\fR=\fIDOUBLE\fR
224 Set the weight of the covariance term in the
229 \fB\-\-nfactor\fR=\fIDOUBLE\fR
230 Set the penalty for non\-compatible sequences in
231 the covariance term of the energy function
235 \fB\-E\fR, \fB\-\-endgaps\fR
236 Score pairs with endgaps same as gap\-gap pairs.
240 \fB\-R\fR, \fB\-\-ribosum_file\fR=\fIribosumfile\fR
241 use specified Ribosum Matrix instead of normal
243 energy model. Matrixes to use should be 6x6
244 matrices, the order of the terms is AU, CG,
247 \fB\-r\fR, \fB\-\-ribosum_scoring\fR
248 use ribosum scoring matrix. The matrix is
249 chosen according to the minimal and maximal
250 pairwise identities of the sequences in the
256 use old energy evaluation, treating gaps as
261 \fB\-P\fR, \fB\-\-paramFile\fR=\fIparamfile\fR
262 Read energy parameters from paramfile, instead
263 of using the default parameter set.
265 A sample parameter file should accompany your distribution.
266 See the RNAlib documentation for details on the file format.
268 \fB\-\-nsp\fR=\fISTRING\fR
269 Allow other pairs in addition to the usual
272 Its argument is a comma separated list of additionally allowed pairs. If the
273 first character is a "\-" then AB will imply that AB and BA are allowed
275 e.g. RNAfold \fB\-nsp\fR \fB\-GA\fR will allow GA and AG pairs. Nonstandard pairs are
276 given 0 stacking energy.
278 \fB\-e\fR, \fB\-\-energyModel\fR=\fIINT\fR
279 Rarely used option to fold sequences from the
280 artificial ABCD... alphabet, where A pairs B,
281 C\-D etc. Use the energy parameters for GC
282 (\fB\-e\fR 1) or AU (\fB\-e\fR 2) pairs.
284 \fB\-\-betaScale\fR=\fIDOUBLE\fR
285 Set the scaling of the Boltzmann factors
288 The argument provided with this option enables to scale the thermodynamic
289 temperature used in the Boltzmann factors independently from the temperature
290 used to scale the individual energy contributions of the loop types. The
291 Boltzmann factors then become exp(\fB\-dG\fR/(kTn*betaScale)) where k is the
292 Boltzmann constant, dG the free energy contribution of the state, T the
293 absolute temperature and n the number of sequences.
297 Sequences are not weighted. If possible, do not mix very similar and dissimilar
298 sequences. Duplicate sequences, for example, can distort the prediction.
301 Ivo L Hofacker, Stephan Bernhart, Ronny Lorenz
303 .I If you use this program in your work you might want to cite:
305 R. Lorenz, S.H. Bernhart, C. Hoener zu Siederdissen, H. Tafer, C. Flamm, P.F. Stadler and I.L. Hofacker (2011),
306 "ViennaRNA Package 2.0",
307 Algorithms for Molecular Biology: 6:26
309 I.L. Hofacker, W. Fontana, P.F. Stadler, S. Bonhoeffer, M. Tacker, P. Schuster (1994),
310 "Fast Folding and Comparison of RNA Secondary Structures",
311 Monatshefte f. Chemie: 125, pp 167-188
314 The algorithm is a variant of the dynamic programming algorithms of M. Zuker and P. Stiegler (mfe)
315 and J.S. McCaskill (pf) adapted for sets of aligned sequences with covariance information.
317 Ivo L. Hofacker, Martin Fekete, and Peter F. Stadler (2002),
318 "Secondary Structure Prediction for Aligned RNA Sequences",
319 J.Mol.Biol.: 319, pp 1059-1066.
321 Stephan H. Bernhart, Ivo L. Hofacker, Sebastian Will, Andreas R. Gruber, and Peter F. Stadler (2008),
322 "RNAalifold: Improved consensus structure prediction for RNA alignments",
323 BMC Bioinformatics: 9, pp 474
326 .I The energy parameters are taken from:
328 D.H. Mathews, M.D. Disney, D. Matthew, J.L. Childs, S.J. Schroeder, J. Susan, M. Zuker, D.H. Turner (2004),
329 "Incorporating chemical modification constraints into a dynamic programming algorithm for prediction of RNA secondary structure",
330 Proc. Natl. Acad. Sci. USA: 101, pp 7287-7292
332 D.H Turner, D.H. Mathews (2009),
333 "NNDB: The nearest neighbor parameter database for predicting stability of nucleic acid secondary structure",
334 Nucleic Acids Research: 38, pp 280-282
336 If in doubt our program is right, nature is at fault.
338 Comments should be sent to rna@tbi.univie.ac.at.
341 The ALIDOT package http://www.tbi.univie.ac.at/RNA/ALIDOT/