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suppressPackageStartupMessages(library(GenomicRanges))
suppressPackageStartupMessages(library(magrittr))
suppressPackageStartupMessages(library(universalmotif))
library(memes)Inputs
FIMO searches input sequences for occurrances of a motif. runFimo() has two required inputs: fasta-format sequences, with optional genomic coordinate headers, and a set of motifs to detect within the input sequences.
Sequence Inputs:
Sequence input to runFimo() can be as a path to a .fasta formatted file, or as a Biostrings::XStringSet object. Unlike other memes functions, runFimo() does not accept a Biostrings::BStringSetList as input. This is to simplify ranged join operations (see joins) on output data.
By default, runFimo() will parse genomic coordinates from sequence entries from the fasta headers. These are generated automatically if using get_sequences() to generate sequences for input from a GRanges object.
data("example_chip_summits", package = "memes")
dm.genome <- BSgenome.Dmelanogaster.UCSC.dm3::BSgenome.Dmelanogaster.UCSC.dm3
# Take 100bp windows around ChIP-seq summits
summit_flank <- example_chip_summits %>%
plyranges::anchor_center() %>%
plyranges::mutate(width = 100)
# Get sequences in peaks as Biostring::BStringSet
sequences <- summit_flank %>%
get_sequence(dm.genome)
# get_sequence includes genomic coordinate as the fasta header name
names(sequences)[1:2]Motif Inputs:
Motif input to runFimo() can be as a path to a .meme formatted file, a list of universalmotif objects, or a singular universalmotif object. runFimo() will not use any of the default search path behavior for a motif database as in runAme() or runTomTom().
e93_motif <- MotifDb::MotifDb %>%
# Query the database for the E93 motif using it's gene name
MotifDb::query("Eip93F") %>%
# Convert from motifdb format to universalmotif format
universalmotif::convert_motifs() %>%
# The result is a list, to simplify the object, return it as a single universalmotif
.[[1]]
# Rename the motif from it's flybase gene number to a more user-friendly name
e93_motif["name"] <- "E93_FlyFactor"Note about default settings
runFimo() is configured to use different default behavior relative to the commandline and MEME-Suite Server versions. By default, runFimo() runs using text mode, which greatly increases speed and allows returning all detected matches to the input motifs. By default runFimo() will add the sequence of the matched region to the output data; however, this operation can be very slow for large sets of regions and can drastically increase the size of the output data. To speed up the operation and decrease data size, set skip_matched_sequence = TRUE. Sequence information can be added back later using add_sequence().
Data integration with join operations
The plyranges package provides an extended framework for performing range-based operations in R. While several of its utilities are useful for range-based analyses, the join_ functions are particularly useful for integrating FIMO results with input peak information. A few common examples are briefly highlighted below:
plyranges::join_overlap_left() can be used to add peak-level metadata to motif position information:
fimo_results_with_peak_info <- fimo_results %>%
plyranges::join_overlap_left(summit_flank)
fimo_results_with_peak_info[1:5]plyranges::intersect_() can be used to simultaneously subset input peaks to the ranges overlapping motif hits while appending motif-level metadata to each overlap.
Identifying matched sequence
When setting skip_match_sequence = TRUE, FIMO does not automatically return the matched sequence within each hit. These sequences can be easily recovered in R using add_sequence() on the FIMO results GRanges object.
fimo_results_with_seq <- fimo_results %>%
plyranges::join_overlap_left(summit_flank) %>%
add_sequence(dm.genome)Returning the sequence of the matched regions can be used to re-derive PWMs from different match categories as follows (here done for different binding categories):
motifs_by_binding <- fimo_results_with_seq %>%
# Split on parameter of interest
split(mcols(.)$peak_binding_description) %>%
# Convert GRangesList to regular list() to use `purrr`
as.list() %>%
# imap passes the list entry as .x and the name of that object to .y
purrr::imap(~{
# Pass the sequence column to create_motif to generate a PCM
create_motif(.x$sequence,
# Append the binding description to the motif name
name = paste0("E93_", .y))
})Motifs from each category can be visualized with universalmotif::view_motifs()
To allow better comparison to the reference motif, we can append it to the list as follows:
motifs_by_binding <- c(
# Add the E93 FlyFactor motif to the list as a reference
list("E93_FlyFactor" = e93_motif),
motifs_by_binding
)Visualizing the motifs as ICMs reveals subtle differences in E93 motif sequence between each category.
Visualizing the results as a position-probability matrix (PPM) does a better job of demonstrating that the primary differences between each category are coming from positions 1-4 in the matched sequences.
Finally, the sequence-level information can be used to visualize all sequences and their contribution to the final PWM using plot_sequence_heatmap.
Importing Data from previous FIMO Runs
importFimo() can be used to import an fimo.tsv file from a previous run on the MEME server or on the commandline. Details for how to save data from the FIMO webserver are below.
Saving data from FIMO Web Server
To download TSV data from the FIMO Server, right-click the FIMO TSV output link and “Save Target As” or “Save Link As” (see example image below), and save as <filename>.tsv. This file can be read using importFimo().
