MotifScan is a precise and easy-use motif discovery tool based on given motifs. To search for candidate motif targtes in given DNA sequences, the program scans them with a window of the same length as the motif, and defined raw motif score of the sequence S in window as the ratio of the probability to observe target sequence S given the motif's Position Weight Matrix (PWM) M and the probability to observe S given the genome background B. For each annotated motif, we modeled the genome background distribution of motif score by randomly sampling the genome for 10^6 times, and defined the targets of this motif as those candidates whose motif score was higher than the cutoff. And the enrichment of each motif was represented by the ratio of motif target densities at input regions compared to random control regions, together with a p-value calculated from hypergeometric distribution. It is noticeable that MotifScan is not a de novo motif discovery tool.


News
  • 2015-04-20 MotifScan_1.0.4 is released.


Download
Latest Release
Version Download
MotifScan 1.0.4 (2015-04-20) MotifScan_1.0.4.tar.gz
Pre-built Motif
Species Genome Downlaod
H. sapiens hg19
M. musculus mm9
A. thaliana tair10
Motif Name List
Group Download
JASPARE CORE All jaspar_2014_all_motif_list.txt
JASPARE CORE Vertebrata jaspar_2014_vertebrates_motif_list.txt
JASPARE CORE Plantae jaspar_2014_plants_motif_list.txt
JASPARE CORE Nematoda jaspar_2014_nematodes_motif_list.txt
JASPARE CORE Insecta jaspar_2014_insects_motif_list.txt
JASPARE CORE Fungi jaspar_2014_fungi_motif_list.txt

Get Started
Obtain MotifScan

Download package from here.

Then extract the package and build from the source:

tar -zxvf MotifScan_*.*.*.tar.gz
cd MotifScan_*.*.*
sudo python setup install

Or use option --prefix to specify the installation position

python setup install --prefix=/home/jiawei

Under this circumstance, you should add your installation directory to PYTHONPATH and bash PATH:

export PYTHONPATH=/home/jiawei/lib/python2.7/dist-packges:$PYTHONPATH
export PATH=/home/jiawei/bin:$PATH

Motif Discovery

Search regions (-p), compiled motif PWM (-m) and genome (-g and -G) are required for a common motifscan task. And the gene annotation is auto-loaded when specifying the genome identifier or you can specify the gene file explicitly via option -t. With all these files prepared, use the following command to perform the scan:

MotifScan –p Example/SL2548_Peaks.bed –m Example/hg19_motif_demo.txt -g hg19 [-G hg19.fa]

-p peak region file in bed format

-m motif information table

-g reference genome identifier

-G genome sequence file

Compiled Motif PWM file (-m) should be organized as follows:

>MA0143.3 Sox2 max_score:9.572 score_cutoff:0.9799
0.513 0.997 0.001 0.997 0.997 0.546 0.001 0.001
0.201 0.001 0.997 0.001 0.001 0.001 0.001 0.001
0.284 0.001 0.001 0.001 0.001 0.001 0.997 0.997
0.001 0.001 0.001 0.001 0.001 0.452 0.001 0.001

Columns are seperated by tab.

MotifScan package has already contains the pre-compiled motif PWMs stemmed from JASPAR. When running the MotifScan command for the first time, a directory named after .MotifScan will be created under your home directory (that is, /home/jiawei/.MotifScan), then built-in gene annotation files will be copied to that directory.

Also, you should specify the genome sequence via option –G for your first time running and the program will format the sequence and store the formatted sequence in /home/jiawei/.MotifScan

Compile Your Own Motif PWM

If you have some motifs (e.g. Example/motif_pwm_demo.txt) that have not be included in our pre-complied motif collection, you need to compile on your own by using the following command.

MotifCompile –M Example/motif_pwm_demo.txt –g hg19 –G hg19.fa

-M motif raw matrix file

-g genome identified

-G genome fasta file

Motif raw matrix file (Example/motif_pwm_demo.txt) should follow the format as below: motif id and motif name are followed by a positive weighted matrix, and columns are seperated by tabs.

>MA0599.1 KLF5
1429 0 0 3477 0 5051 0 0 0 3915
2023 11900 12008 9569 13611 0 13611 13611 13135 5595
7572 0 0 0 0 5182 0 0 0 0
2587 1711 1603 565 0 3378 0 0 476 4101

Five motif files representing different score cutoff are generated.


Utility (Under Construction)
Here, we provide MotifScan Lite, the online version of MotifScan with trimmed components.
or


Manual
MotifScan

MotifScan is generally performed by 3 steps:

Step 1: motif discovery on input regions
Step 2: motif discovery on random control regions
Step 3: find out the enriched motifs of input regions compared to random control regions

For the fisrt time running MotifScan:
A common MotifScan procedure requires several data files (motif information, genome information and gene information), motif information and gene information are included in our packages. When you run the motifscan on the genome for the first time, a directory (.MotifScan) that storing above information will be created in your home directory. Under that directory, all the pre-compiled motifs and gene annotation are contained except the genome information.

Thus, you should specify the genome sequence file (.fa or .fasta) via option -G when you run the MotifScan for the first time to compile the genome information. After that, if you want to do MotifScan again on this genome, option -G can be omitted because the genome information has been stored under the .MotifScan directory.

Take hg19 as an example, after your initial motif scanning, gene annotation is located under .MotifScan/gene/hg19/ and genome information is located under .MotifScan/genome/hg19/.

Command Line

MotifScan -p Example/SL2548_Peaks.bed -m Example/motif_list_example.txt -g hg19 [-G hg19.fa]

Note: -G option is required if you run the command on hg19 for the first time.

Options

-p region file.

MotifScan is occurred in the regions specified in this file. It is a customized bed format file. bed5col is default format, but you can also assign your region file format by option -f (see option -f for details)

-g genome identifier.

Please use UCSC genome identifier if possible. Now MotifScan pre-compiled the following genome, hg19, mm9, tair10. You can specify other genomes by command MotifCompile

-m compiled motif PWM.

Motif matrix file including pvalue cutoff and perfect ratio.

-G genonme sequence.

If the genome information is not provided, you should specify the genome sequence file (.fa) by this option.

-f region file format.

Default: bed5col

MotifScan supports region files in 5 formats:

1. bed3col: a 3-column bed file, representing chromosome, start and end respecteively. It the simplest format containing least information.

e.g.
chr1 1711300 1711875
chr1 2179623 2180121
chr1 4717268 4717738
chr1 5285608 5286259
chr1 5327637 5327962

2. bed4col: a 4-column bed file, one more column representing region summit than bed3col. The summit value is the relative distance to the start site.

e.g.
chr1 1711300 1711875 288
chr1 2179623 2180121 249
chr1 4717268 4717738 235
chr1 5285608 5286259 326
chr1 5327637 5327962 163

3. bed5col: a 5-column bed file, one more column representing some statistics (like pvalue in MACS or mvalue in MAnorm) than bed4col.

e.g.
chr1 1711300 1711875 288 442.69
chr1 2179623 2180121 249 118.61
chr1 4717268 4717738 235 137.59
chr1 5285608 5286259 326 238.79
chr1 5327637 5327962 163 224.29

4. manorm: it is a manorm file usually with mvalue column.

e.g.
chr start end summit MAnorm Mvalue MAnorm Avalue MAnorm Pvalue common or unique
chrY 2653916 2656315 1172 6.76 2.38 2.05e-15 H1hesc H3k4me3 Bro
chrY 2709282 2711375 683 7.30 2.65 8.93e-23 H1hesc H3k4me3 Bro
chrY 2801707 2804982 2024 7.15 2.57 2.05e-20 H1hesc H3k4me3 Bro

5. macs: it is peak file called by MACS, usually with pvalue column.

e.g.
chr start end length summit tags -10*log10(pvalue) fold enrichment FDR
chr1 27644 30246 2603 1257 375 1609.14 19.29 0.14
chr1 540240 541130 891 516 24 59.25 6.12 10.98
chr1 712921 715877 2957 973 384 2813.05 46.96 0.10
chr1 761379 763880 2502 1094 332 2005.89 34.28 0.14
chr1 838832 839681 850 639 24 62.84 6.68 9.29

Basically, they are all bed files and bed3col, bed4col, bed5col format does not have the header line.

-l motif_list.

If specified, the program will perform scanning only on the given motifs. Otherwise, MotifScan will be performed on each motif provided in the compiled motif PWM file (specified by option -m). The motif list should be a subset of the available motifs from compiled motif PWM file, and the file should be organized as follows:

e.g.
Pou5f1
SOX10
ARID3A
Prrx2
Sox5
NFIC
Sox2
POU2F2
TEAD1
Sox3
---EOF---

--o output directory.

Specify the output directory. If the directory is not exites, program will create it. The default output value would be motifscan_output_[Prefix_of_input_region_file]

Advanced Options
[enrichment analysis options]

-e enrichment flag.

With this flag on, MotifScan program will only perform step 1, that is not scanning on the random control regions and not performing enrichment analysis.

-s target site flag.

With this flag on, MotifScan will report target sequence and absolute position of each motif

-t gene annotation file.

Gene annotation is meaningful for generating random controls in step 2.

If all regions are in the promoter region, then we can picked up random regions just from the gene promoters instead of the whole genome. And it is believed that sequence characters on promoters are very differnt from those on whole genome. Thus, the gene annotation is an indispensable information. However, in most cases, you do not need really give the gene annotation file explicitly. When you specify the genome name via option -g, the corresponding gene annotation is automatically loaded unless you running MotifScan on your custom genome.

--random_times.

The number of times that picking up the random regions depends on the number f regions you provided. By default, it is 5 times. For example, you provided 1000 regions, then 5000 random regions will be generated.

[search region options]

-r

possible values: genome, promoter, distal
genome: perform MotifScan on the all regions, the default.
promoter: perform MotifScan on regions that only positioned at promoters
distal: perform MotifScan on regions that only positioned at distals

--promoter_up

Only meaningful when option -r is promoter or distal. Define promoter regions: upstream distance to TSS. Default: 4000

--promoter_down

Only meaningful when option -r is promoter or distal. Define promoter regions: downstream distance to TSS. Default: 2000

--region_length

The region length around region summit. If summit is not provided in the region file, region midpoint is viewed as the summit. Default:1000

Result Interpretation
motif_enrichment.csv

All analyzed motifs are listed and sorted by enrichment p-value in the ascending order.

e.g.

name target number rnd target number fold change enrich pvalue deplete pvalue pvalue corrected
TAL1::GATA1 274 1214 1.12 0.033 0.971 0.067
TAL1::TCF3 2691335 1.007 0.464 0.562 0.929

peak_motif_score.csv

The table can be divided into two parts, the first 5 columns are the region information part which briefly derived from the region file that user specified and the second part is the motif score information. Each motif has a score measuring the binding affinity for each region sequence.

e.g.

chrstartendsummitvalueZNF263.ratioGATA1.ratio ...
chr134724923473222347275264.520.830.34 ...
chr134733903474130 347400554.190.730.07...
chr147740184774765477457852.820.84 0.62...

motif score is the measurement of the probability of the motif occurrence, that is the binding affinity of this motif on the site.

peak_motif_tarnum.csv

It is a also detail information table for each region’s motif target number for each motif. The file structure is similar to the peak_motif_score.csv, except the bold font represents the motif target number instead of the motif score.

e.g.

chr start end summit value ZNF263.tarnum GATA1.tarnum...
chr1 3472492 34732223472752 64.52 3 0...
chr1 3473390 34741303474005 54.19 2 0...
chr1 4774018 47747654774578 52.82 0 1...
motif target sites/*

Only appears when option -s is on. The directory contains all the motif target site information of all candidate motifs. Each motif forms an independent file that named after [motif_name]_target_site.txt. The fisrt 3 columns are the motif target site coordinate on the genome. The 4th column is the corresponding target sequence and the motif score of the this motif occurrence is indicated in the last column

e.g. The motif target site of TAL1::GATA1

chrstartendsequencemotif score
chr12368418223684200ATTATCATTTCAGTGCAC0.57
chr13486164434861662CTGTTTCTGGAAGATATT0.54
chr13602069336020711CTGCATTCAACAGATACT0.56
chr13851932938519347CAGGATCAAGCTGATAAA0.54
chr13941969339419711CTTATCTTCTATCAACAG0.73
chr15330185953301877TTTATCTTCTAGTCCCAC0.58
chr16419727264197290CCTATCACGGGAGAACAG0.75
chr16493876964938787TTTATCTCCTCCCAACTG0.56
plot/*

Under this directory, graphs for each motif will be generated.

MotifCompile

You can use MotifCompile to build your own motif and genome data.
MotifCompile extracted 1 million regions from whole genome, and calculated motif score on for each motif on each region, this step is called simulation, which is usually time-consuming. Meanwhile, each motif's raw matrix are normalized, consensus sequence are generated, genome background and chromosome are calculated, which are essential information for our MotifScan task.

Command Line

MotifCompile -M jaspar_ver.txt -g hg19 [-G hg19.fa]

4.2 Options

-G genome sequence file.

The option can be omitted when the genome has been built previously.

-g genome identifier.

If the genome name already exists in the MotifScan data set. MotifCompile will try to find the associated genome sequence automatically. Thus, above option -G can be omitted in this case.

-M motif PWM file.

The matrix should follow the JASPAR raw motif matrix format, that is: each motif id followed by a motif name (separated by tab), they are in a line started by '>' and the matrix is like this:

3976 1495 0 13819 0 0 0 0 10453 0 2184
2073 4490 13819 0 4863 0 0 0 745 9929 6633
7125 1909 0 0 1370 13819 0 13819 2568 729 455
645 5925 0 0 7586 0 13819 0 53 3161 4547

4 rows represent A, C, G, T while columns represent the base positions, elements are separated by tab.

News


  • 2015-04-20: MotifScan_1.0.4 is released.

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