In ddiez/motiftools: Tools to analyze sequence motifs

library(BiocStyle)
library(ggplot2)
library(ggthemes)
theme_set(theme_base(base_size = 12) + theme(plot.background = element_rect(color = NA)))

Introduction

motiftools is a package for the analysis and interpretation of sequence motifs. Its initial implementation is focused on protein motif analysis obtained from the MEME suite, but its functionality may extend to other applications and tools, including analysis of nucleotide sequences and motif analysis tools found in Bioconductor or CRAN.

Motif ranges are stored as RangedData objects (r Biocpkg("IRanges")) package). This allows to include metadata, like p-value, score and other output from the search process.

library(motiftools)
library(ggseqlogo)
library(Biostrings)
library(ggtree)

Plotting functions

Alignments

You can align two protein sequences specifying a particular substitution matrix (e.g. BLOSUM62).

data("BLOSUM62", package = "Biostrings")
align("EGGALAPA", "DGGIVLPSAV", score.matrix = BLOSUM62, type = "global")$alignment
align("EGGALAPA", "DGGIVFFRF", score.matrix = BLOSUM62, type = "local")$alignment

Motifs

# load MEME results.
f <- system.file("files/meme.xml", package = "motiftools")
x_meme <- readMEME(f)
m_meme <- getMotifMatrix(x_meme)
plotMotifMatrix(m_meme, fill = c("white", "grey"))

# load FIMO results.
f <- system.file("files/fimo.xml", package = "motiftools")
x_fimo <- readFIMO(f)
m_fimo <- getMotifMatrix(x_fimo)
plotMotifMatrix(list(meme = m_meme, fimo = m_fimo), fill = c("white", "grey"), col = "black")

# load MAST results.
f <- system.file("files/mast.xml", package = "motiftools")
x_mast <- readMAST(f)
m_mast <- getMotifMatrix(x_mast)
plotMotifMatrix(list("MEME motifs" = m_meme, "FIMO motifs" = m_fimo, "MAST motifs" = m_mast), fill = c("white", "grey"), col = "black")

# highlight some sequences.
annot <- matrix(1, nrow = nseq(x_meme), ncol = 1, dimnames = list(sequenceNames(x_meme)))
annot["RASH_MOUSE", ] <- 2
annot["RASK_MOUSE", ] <- 3
annot
plotMotifMatrix(list("MEME motifs" = m_meme, "FIMO motifs" = m_fimo, "MAST motifs" = m_mast), fill = c("white", "grey"), col = "black", annot = list(annot), annot.fill = list(c("white", "red", "blue")), show.tips = TRUE)

# plot a motif model logo (from MEME output).
p <- pwm(x_meme)[["motif_4"]]
ggseqlogo(p)

You can compare the motifs obtained with MEME using the TOMTOM tool. Then you can read the results and look at the comparison matrix.

f <- system.file("files/tomtom.xml", package = "motiftools")
x_tomtom <- readTOMTOM(f)
plotMotifMatches(x_tomtom)
plotMotifMatches(x_tomtom, fill = "e_value", color = "black")

Architectures

Sometimes different motifs are arranged in different order in different sequences, suggesting processes of reordering. Motif ordering is called in motiftools architectures. You can obtain the architectures from the sequences used to compute the MEME motifs and compute the similarity between them.

getMotifsBySeq(x_meme) # returns a list with the ordered motif composition as a character vector.
getMotifArchString(x_meme) # returns a list with a single string with the architecture encoded.

# compute sequence motif Jaccard similarity.
s <- getMotifSimilarity(x_meme)
s
plotMotifArchSimilarity(s)

# compute sequence motif architecture Jaccard similarity.
s <- getMotifArchSimilarity(x_meme)
s
plotMotifArchSimilarity(s)

Sequences

We can visualize MSA conservation together with a phylogenetic tree.

ras_tree <- read.tree(system.file("files/ras.tree", package = "motiftools"))
ras_aln <- readAAMultipleAlignment(system.file("files/ras.faln", package = "motiftools"))
rownames(ras_aln) <- sub(" .*", "", rownames(ras_aln)) # need to fix sequence names.
plotConservationMatrix(ras_aln, ras_tree, show.tips = TRUE)