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bioconductor-tfbstools

TFBSTools is a package for the analysis and manipulation of transcription factor binding sites. It includes matrices conversion between Position Frequency Matirx (PFM), Position Weight Matirx (PWM) and Information Content Matrix (ICM). It c

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TFBSTools

Dependencies & Environment

Package-intrinsic requirements from the Bioconductor landing page — reproduce in any R environment.

  • Version: 1.50.0 · Bioconductor: 3.23 · R: ≥ 4.6
  • Imports: Biobase, Biostrings, pwalign, BiocGenerics, BiocParallel, BSgenome, caTools, DirichletMultinomial, Seqinfo, GenomicRanges, gtools, IRanges, DBI, RSQLite, rtracklayer, seqLogo, S4Vectors, TFMPvalue, XML, XVector
  • Install: BiocManager::install("TFBSTools")

When to Use

  • Matrix Conversion: Converting Position Frequency Matrices (PFMs) to Position Weight Matrices (PWMs) or Information Content Matrices (ICMs) using toPWM and toICM.
  • Motif Scanning: Scanning nucleotide sequences or pairwise alignments for transcription factor binding sites using searchSeq or searchAln.
  • Database Querying: Querying and retrieving matrix data from the JASPAR database using getMatrixSet.
  • Random Profile Generation: Generating random profile matrices via permutation or Dirichlet multinomial mixture models using permuteMatrix and rPWMDmm.
  • Visualization: Visualizing sequence logos for basic PWMs or Transcription Factor Flexible Models (TFFMs) using seqLogo.

When NOT to Use

  • For high-throughput scanning of thousands of large genomic regions, use motifmatchr instead, because searchSeq is optimized for targeted DNAString or DNAStringSet objects rather than massive GRanges peak sets.
  • For de novo motif discovery without external dependencies, use universalmotif because runMEME requires the external MEME suite to be installed on the system.

Data Requirements

  • Input Format: PFMatrix objects, DNAString or DNAStringSet for sequences, and Axt objects for alignments.
  • Structure: 4-row matrices representing A, C, G, T counts or probabilities.
  • Normalization State: Raw position frequency counts for PFMs, which are then converted to log2 probability ratios for PWMs.

Key Parameters

  • type (default): The type of matrix conversion in toPWM (e.g., "log2probratio") or the type of p-value calculation in pvalues (e.g., "TFMPvalue").
  • pseudocounts (default): Numeric value (default 0.8) added to correct small counts or eliminate zero values before log transformation in toPWM.
  • bg (default): A named numeric vector representing background nucleotide frequencies (e.g., c(A=0.25, C=0.25, G=0.25, T=0.25)).
  • min.score (default): Minimum score threshold (e.g., "60%" or "80%") for a sequence match to be reported in searchSeq or searchAln.
  • schneider (default): Logical in toICM indicating whether to apply the Schneider correction.
  • strand (default): Controls which strand is searched in searchSeq (e.g., "*" for both strands).

Best Practices

  • Use pseudocounts=0.8 in toPWM rather than the square root of sequences to avoid overly harsh corrections on zero values.
  • When scanning sequences with searchSeq, use pvalues with type="TFMPvalue" to calculate empirical p-values for the match scores.
  • Use writeGFF3 or writeGFF2 to export SiteSet objects into standard genomic formats for downstream visualization.

Common Pitfalls

  • Pitfall: runMEME fails to execute.
    Fix: Ensure the external MEME software suite is installed and the binary argument correctly points to the executable.
  • Pitfall: searchSeq returns too many false positive hits.
    Fix: Increase the min.score threshold (e.g., from "60%" to "90%") to enforce stricter matching.
  • Pitfall: toPWM throws an error due to zero counts.
    Fix: Ensure pseudocounts is set to a positive value (like 0.8) to eliminate zero values before log transformation.

Alternatives

  • motifmatchr: For faster, parallelized motif scanning across large GRanges objects.
  • universalmotif: For comprehensive motif manipulation and de novo discovery without relying strictly on external binaries like MEME.
  • Biostrings: For basic PWM matching (matchPWM), though it lacks the advanced statistical models and TFFM support of TFBSTools.

Citations

  • Tan G, Lenhard B (2016). TFBSTools: an R/Bioconductor package for transcription factor binding site analysis. Bioinformatics.
  • Mathelier and Wasserman (2013). The next generation of transcription factor binding site, TFFM.

References

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