title: Benchmarking the next generation of homology inference tools.
abstract: Over the last decades, vast numbers of sequences were deposited in public databases. Bioinformatics tools allow homology and consequently functional inference for these sequences. New profile-based homology search tools have been introduced, allowing reliable detection of remote homologs, but have not been systematically benchmarked. To provide such a comparison, which can guide bioinformatics workflows, we extend and apply our previously developed benchmark approach to evaluate the 'next generation' of profile-based approaches, including CS-BLAST, HHSEARCH and PHMMER, in comparison with the non-profile based search tools NCBI-BLAST, USEARCH, UBLAST and FASTA.
  
  
  
  We generated challenging benchmark datasets based on protein domain architectures within either the PFAM + Clan, SCOP/Superfamily or CATH/Gene3D domain definition schemes. From each dataset, homologous and non-homologous protein pairs were aligned using each tool, and standard performance metrics calculated. We further measured congruence of domain architecture assignments in the three domain databases.
  
  
  
  CSBLAST and PHMMER had overall highest accuracy. FASTA, UBLAST and USEARCH showed large trade-offs of accuracy for speed optimization.
  
  
  
  Profile methods are superior at inferring remote homologs but the difference in accuracy between methods is relatively small. PHMMER and CSBLAST stand out with the highest accuracy, yet still at a reasonable computational cost. Additionally, we show that less than 0.1% of Swiss-Prot protein pairs considered homologous by one database are considered non-homologous by another, implying that these classifications represent equivalent underlying biological phenomena, differing mostly in coverage and granularity.
  
  
  
  Benchmark datasets and all scripts are placed at (http://sonnhammer.org/download/Homology_benchmark).
  
  
  
  forslund@embl.de
  
  
  
  Supplementary data are available at Bioinformatics online.
authors.affiliation: Science for Life Laboratory, Stockholm Bioinformatics Center, Department of Biochemistry and Biophysics, Stockholm University, Stockholm SE-10691, Sweden.
authors.forename: Ganapathi Varma
authors.initials: GV
authors.lastname: Saripella
authors.affiliation: Science for Life Laboratory, Stockholm Bioinformatics Center, Department of Biochemistry and Biophysics, Stockholm University, Stockholm SE-10691, Sweden.
authors.forename: Erik L L
authors.initials: EL
authors.lastname: Sonnhammer
authors.affiliation: European Molecular Biology Laboratory, Structural and Computational Biology Unit, Heidelberg 69117, Germany.
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Forslund
mesh_terms: D019985:Benchmarking; D030562:Databases, Protein; D059014:High-Throughput Nucleotide Sequencing; D011506:Proteins; D017385:Sequence Homology; D017386:Sequence Homology, Amino Acid
chemical_list: D011506:Proteins