Ubuntu Manpage: RNAplfold - manual page for RNAplfold 2.6.4

Provided by: vienna-rna_2.6.4+dfsg-1build2_amd64

NAME

       RNAplfold - manual page for RNAplfold 2.6.4

SYNOPSIS

       RNAplfold [OPTION]...

DESCRIPTION

RNAplfold 2.6.4

calculate locally stable secondary structure - pair probabilities

Computes local pair probabilities for base pairs with a maximal span of L. The probabilities are averaged
over all windows of size L that contain the base pair. For a sequence of length n and a window size of L
the algorithm uses only O(n+L*L) memory and O(n*L*L) CPU time. Thus it is practical to "scan" very large
genomes for short stable RNA structures.

Output consists of a dot plot in postscript file, where the averaged pair probabilities can easily be
parsed and visually inspected.

The -u option makes i possible to compute the probability that a stretch of x consequtive nucleotides is
unpaired, which is useful for predicting possible binding sites. Again this probability is averaged over
all windows containing the region.

WARNING! Output format changed!!

The output is a plain text matrix containing on each line a position i followed by the probability that i
is unpaired, [i-1..i] is unpaired [i-2..i] is unpaired and so on to the probability that [i-x+1..i] is
unpaired.

-h, --help
Print help and exit

--detailed-help
Print help, including all details and hidden options, and exit

--full-help
Print help, including hidden options, and exit

-V, --version
Print version and exit

-v, --verbose
Be verbose.

(default=off)

I/O Options:
Command line options for input and output (pre-)processing

-c, --cutoff=FLOAT
Report only base pairs with an average probability larger than 'cutoff' in the dot plot.

(default=`0.01')

-o, --print_onthefly
Save memory by printing out everything during computation.

(default=off)

NOTE: activated per default for sequences over 1M bp.

-O, --opening_energies
Switch output from probabilities to their logarithms.

(default=off)

This is NOT exactly the mean energies needed to unfold the respective stretch of bases! (implies
--ulength option).

--plex_output
Create additional output files for RNAplex.

(default=off)

-b, --binaries
Output accessibility profiles in binary format. (default=off)

The binary files produced by RNAplfold do not need to be parsed by RNAplex,

so that they are directly loaded into memory. This is useful when large sequences have to be
searched for putative hybridization sites. Another advantage of the binary format is the 50% file
size decrease.

--noconv
Do not automatically substitute nucleotide "T" with "U".

(default=off)

--auto-id
Automatically generate an ID for each sequence. (default=off)

The default mode of RNAplfold is to automatically determine an ID from the input sequence data if
the input file format allows to do that. Sequence IDs are usually given in the FASTA header of
input sequences. If this flag is active, RNAplfold ignores any IDs retrieved from the input and
automatically generates an ID for each sequence. This ID consists of a prefix and an increasing
number. This flag can also be used to add a FASTA header to the output even if the input has none.

--id-prefix=STRING
Prefix for automatically generated IDs (as used in output file names).

(default=`sequence')

If this parameter is set, each sequences' FASTA id will be prefixed with the provided string.
FASTA ids then take the form ">prefix_xxxx" where xxxx is the sequence number. Hence, the output
files will obey the following naming scheme: "prefix_xxxx_dp.ps" (dot-plot), "prefix_xxxx_lunp"
(unpaired probabilities), etc. Note: Setting this parameter implies --auto-id.

--id-delim=CHAR
Change the delimiter between prefix and increasing number for automatically generated IDs (as used
in output file names).

(default=`_')

This parameter can be used to change the default delimiter "_" between the prefix string and the
increasing number for automatically generated ID.

--id-digits=INT
Specify the number of digits of the counter in automatically generated alignment IDs.

(default=`4')

When alignments IDs are automatically generated, they receive an increasing number, starting with
1. This number will always be left-padded by leading zeros, such that the number takes up a
certain width. Using this parameter, the width can be specified to the users need. We allow
numbers in the range [1:18]. This option implies --auto-id.

--id-start=LONG
Specify the first number in automatically generated IDs.

(default=`1')

When sequence IDs are automatically generated, they receive an increasing number, usually starting
with 1. Using this parameter, the first number can be specified to the users requirements. Note:
negative numbers are not allowed. Note: Setting this parameter implies to ignore any IDs
retrieved from the input data, i.e. it activates the --auto-id flag.

--filename-delim=CHAR
Change the delimiting character used in sanitized filenames.

(default=`ID-delimiter')

This parameter can be used to change the delimiting character used while sanitizing filenames,
i.e. replacing invalid characters. Note, that the default delimiter ALWAYS is the first character
of the "ID delimiter" as supplied through the --id-delim option. If the delimiter is a whitespace
character or empty, invalid characters will be simply removed rather than substituted. Currently,
we regard the following characters as illegal for use in filenames: backslash '\', slash '/',
question mark '?', percent sign '%', asterisk '*', colon ':', pipe symbol '|', double quote '"',
triangular brackets '<' and '>'.

--filename-full
Use full FASTA header to create filenames. (default=off)

This parameter can be used to deactivate the default behavior of limiting output filenames to the
first word of the sequence ID. Consider the following example: An input with FASTA header
'>NM_0001 Homo Sapiens some gene' usually produces output files with the prefix "NM_0001" without
the additional data available in the FASTA header, e.g. "NM_0001_ss.ps" for secondary structure
plots. With this flag set, no truncation of the output filenames is done, i.e. output filenames
receive the full FASTA header data as prefixes. Note, however, that invalid characters (such as
whitespace) will be substituted by a delimiting character or simply removed, (see also the
parameter option --filename-delim).

Algorithms:
Select and change parameters of (additional) algorithms which should be included in the
calculations.

-W, --winsize=size
Average the pair probabilities over windows of given size.

(default=`70')

-L, --span=size
Set the maximum allowed separation of a base pair to span.

By setting the maximum base pair span no pairs (i,j) with j-i > span will be allowed. Defaults to
winsize if parameter is omitted.

-u, --ulength=length
Compute the mean probability that regions of length 1 to a given length are unpaired.

(default=`31')

Output is saved in a '_lunp' file.

--betaScale=DOUBLE
Set the scaling of the Boltzmann factors. (default=`1.')

The argument provided with this option is used to scale the thermodynamic temperature in the
Boltzmann factors independently from the temperature of the individual loop energy contributions.
The Boltzmann factors then become 'exp(- dG/(kT*betaScale))' where 'k' is the Boltzmann constant,
'dG' the free energy contribution of the state and 'T' the absolute temperature.

-S, --pfScale=DOUBLE
In the calculation of the pf use scale*mfe as an estimate for the ensemble free energy (used to
avoid overflows).

(default=`1.07')

The default is 1.07, useful values are 1.0 to 1.2. Occasionally needed for long sequences.

Structure Constraints:
Command line options to interact with the structure constraints feature of this program

--shape=filename
Use SHAPE reactivity data to guide structure predictions.

--shapeMethod=method
Select SHAPE reactivity data incorporation strategy.

(default=`D')

The following methods can be used to convert SHAPE reactivities into pseudo energy contributions.

'D': Convert by using the linear equation according to Deigan et al 2009.

Derived pseudo energy terms will be applied for every nucleotide involved in a stacked pair. This
method is recognized by a capital 'D' in the provided parameter, i.e.: --shapeMethod="D" is the
default setting. The slope 'm' and the intercept 'b' can be set to a non-default value if
necessary, otherwise m=1.8 and b=-0.6. To alter these parameters, e.g. m=1.9 and b=-0.7, use a
parameter string like this: --shapeMethod="Dm1.9b-0.7". You may also provide only one of the two
parameters like: --shapeMethod="Dm1.9" or --shapeMethod="Db-0.7".

'Z': Convert SHAPE reactivities to pseudo energies according to Zarringhalam

et al 2012. SHAPE reactivities will be converted to pairing probabilities by using linear mapping.
Aberration from the observed pairing probabilities will be penalized during the folding recursion.
The magnitude of the penalties can affected by adjusting the factor beta (e.g.
--shapeMethod="Zb0.8").

'W': Apply a given vector of perturbation energies to unpaired nucleotides

according to Washietl et al 2012. Perturbation vectors can be calculated by using RNApvmin.

--shapeConversion=method
Select method for SHAPE reactivity conversion.

(default=`O')

This parameter is useful when dealing with the SHAPE incorporation according to Zarringhalam et
al. The following methods can be used to convert SHAPE reactivities into the probability for a
certain nucleotide to be unpaired.

'M': Use linear mapping according to Zarringhalam et al. 'C': Use a cutoff-approach to divide
into paired and unpaired nucleotides (e.g. "C0.25") 'S': Skip the normalizing step since the input
data already represents probabilities for being unpaired rather than raw reactivity values 'L':
Use a linear model to convert the reactivity into a probability for being unpaired (e.g.
"Ls0.68i0.2" to use a slope of 0.68 and an intercept of 0.2) 'O': Use a linear model to convert
the log of the reactivity into a probability for being unpaired (e.g. "Os1.6i-2.29" to use a slope
of 1.6 and an intercept of -2.29)

--commands=filename
Read additional commands from file

Commands include hard and soft constraints, but also structure motifs in hairpin and interior
loops that need to be treeted differently. Furthermore, commands can be set for unstructured and
structured domains.

Energy Parameters:
Energy parameter sets can be adapted or loaded from user-provided input files

-T, --temp=DOUBLE
Rescale energy parameters to a temperature of temp C. Default is 37C.

(default=`37.0')

-P, --paramFile=paramfile
Read energy parameters from paramfile, instead of using the default parameter set.

Different sets of energy parameters for RNA and DNA should accompany your distribution. See the
RNAlib documentation for details on the file format. The placeholder file name 'DNA' can be used
to load DNA parameters without the need to actually specify any input file.

-4, --noTetra
Do not include special tabulated stabilizing energies for tri-, tetra- and hexaloop hairpins.

(default=off)

Mostly for testing.

--salt=DOUBLE
Set salt concentration in molar (M). Default is 1.021M.

-m, --modifications[=STRING]
Allow for modified bases within the RNA sequence string.

(default=`7I6P9D')

Treat modified bases within the RNA sequence differently, i.e. use corresponding energy
corrections and/or pairing partner rules if available. For that, the modified bases in the input
sequence must be marked by their corresponding one-letter code. If no additional arguments are
supplied, all available corrections are performed. Otherwise, the user may limit the modifications
to a particular subset of modifications, resp. one-letter codes, e.g. -mP6 will only correct for
pseudouridine and m6A bases.

Currently supported one-letter codes and energy corrections are:

'7': 7-deaza-adenonsine (7DA)

'I': Inosine

'6': N6-methyladenosine (m6A)

'P': Pseudouridine

'9': Purine (a.k.a. nebularine)

'D': Dihydrouridine

--mod-file=STRING
Use additional modified base data from JSON file.

Model Details:
Tweak the energy model and pairing rules additionally using the following parameters

-d, --dangles=INT
Specify "dangling end" model for bases adjacent to helices in free ends and multi-loops.

(default=`2')

With -d2 dangling energies will be added for the bases adjacent to a helix on both sides in any
case while -d0 ignores dangling ends altogether (mostly for debugging).

--noLP Produce structures without lonely pairs (helices of length 1).

(default=off)

For partition function folding this only disallows pairs that can only occur isolated. Other pairs
may still occasionally occur as helices of length 1.

--noGU Do not allow GU pairs.

(default=off)

--noClosingGU
Do not allow GU pairs at the end of helices.

(default=off)

--nsp=STRING
Allow other pairs in addition to the usual AU,GC,and GU pairs.

Its argument is a comma separated list of additionally allowed pairs. If the first character is a
"-" then AB will imply that AB and BA are allowed pairs, e.g. --nsp="-GA" will allow GA and AG
pairs. Nonstandard pairs are given 0 stacking energy.

-e, --energyModel=INT
Set energy model.

Rarely used option to fold sequences from the artificial ABCD... alphabet, where A pairs B, C-D
etc. Use the energy parameters for GC (-e 1) or AU (-e 2) pairs.

--helical-rise=FLOAT
Set the helical rise of the helix in units of Angstrom.

(default=`2.8')

Use with caution! This value will be re-set automatically to 3.4 in case DNA parameters are loaded
via -P DNA and no further value is provided.

--backbone-length=FLOAT
Set the average backbone length for looped regions in units of Angstrom.

(default=`6.0')

Use with caution! This value will be re-set automatically to 6.76 in case DNA parameters are
loaded via -P DNA and no further value is provided.

REFERENCES

       If you use this program in your work you might want to cite:

       R.  Lorenz,  S.H. Bernhart, C. Hoener zu Siederdissen, H. Tafer, C. Flamm, P.F. Stadler and I.L. Hofacker
       (2011), "ViennaRNA Package 2.0", Algorithms for Molecular Biology: 6:26

       I.L. Hofacker, W. Fontana, P.F. Stadler, S. Bonhoeffer, M. Tacker, P. Schuster (1994), "Fast Folding  and
       Comparison of RNA Secondary Structures", Monatshefte f. Chemie: 125, pp 167-188

       R.  Lorenz,  I.L. Hofacker, P.F. Stadler (2016), "RNA folding with hard and soft constraints", Algorithms
       for Molecular Biology 11:1 pp 1-13

       S. H. Bernhart, U. Mueckstein, and I.L. Hofacker (2011), "RNA Accessibility in  cubic  time",  Algorithms
       Mol Biol. 6: 3.

       S. H. Bernhart, I.L. Hofacker, and P.F. Stadler (2006), "Local Base Pairing Probabilities in Large RNAs",
       Bioinformatics: 22, pp 614-615

       A.F.  Bompfuenewerer, R. Backofen, S.H. Bernhart, J. Hertel, I.L. Hofacker, P.F. Stadler, S. Will (2007),
       "Variations on RNA Folding and Alignment: Lessons from Benasque", J. Math. Biol.

       The energy parameters are taken from:

       D.H. Mathews, M.D. Disney, D. Matthew, J.L. Childs, S.J. Schroeder,  J.  Susan,  M.  Zuker,  D.H.  Turner
       (2004),  "Incorporating  chemical  modification  constraints  into  a  dynamic  programming algorithm for
       prediction of RNA secondary structure", Proc. Natl. Acad. Sci. USA: 101, pp 7287-7292

       D.H Turner, D.H. Mathews (2009), "NNDB: The nearest neighbor parameter database for predicting  stability
       of nucleic acid secondary structure", Nucleic Acids Research: 38, pp 280-282

AUTHOR

       Stephan H Bernhart, Ivo L Hofacker, Peter F Stadler, Ronny Lorenz

REPORTING BUGS

       If in doubt our program is right, nature is at fault.  Comments should be sent to rna@tbi.univie.ac.at.

NAME

SYNOPSIS

DESCRIPTION

REFERENCES

AUTHOR

REPORTING BUGS

SEE ALSO