Fast Statistical Alignment

We describe a new program for the alignment of multiple biological sequences that is both statistically motivated and fast enough for problem sizes that arise in practice. Our Fast Statistical Alignment program is based on pair hidden Markov models which approximate an insertion/deletion process on a tree and uses a sequence annealing algorithm to combine the posterior probabilities estimated from these models into a multiple alignment. FSA uses its explicit statistical model to produce multiple alignments which are accompanied by estimates of the alignment accuracy and uncertainty for every column and character of the alignment—previously available only with alignment programs which use computationally-expensive Markov Chain Monte Carlo approaches—yet can align thousands of long sequences. Moreover, FSA utilizes an unsupervised query-specific learning procedure for parameter estimation which leads to improved accuracy on benchmark reference alignments in comparison to existing programs. The centroid alignment approach taken by FSA, in combination with its learning procedure, drastically reduces the amount of false-positive alignment on biological data in comparison to that given by other methods. The FSA program and a companion visualization tool for exploring uncertainty in alignments can be used via a web interface at http://orangutan.math.berkeley.edu/fsa/, and the source code is available at http://fsa.sourceforge.net/.Author Summary: Biological sequence alignment is one of the fundamental problems in comparative genomics, yet it remains unsolved. Over sixty sequence alignment programs are listed on Wikipedia, and many new programs are published every year. However, many popular programs suffer from pathologies such as aligning unrelated sequences and producing discordant alignments in protein (amino acid) and codon (nucleotide) space, casting doubt on the accuracy of the inferred alignments. Inaccurate alignments can introduce large and unknown systematic biases into downstream analyses such as phylogenetic tree reconstruction and substitution rate estimation. We describe a new program for multiple sequence alignment which can align protein, RNA and DNA sequence and improves on the accuracy of existing approaches on benchmarks of protein and RNA structural alignments and simulated mammalian and fly genomic alignments. Our approach, which seeks to find the alignment which is closest to the truth under our statistical model, leaves unrelated sequences largely unaligned and produces concordant alignments in protein and codon space. It is fast enough for difficult problems such as aligning orthologous genomic regions or aligning hundreds or thousands of proteins. It furthermore has a companion GUI for visualizing the estimated alignment reliability.