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Updated: Aug 6, 2026

Demonstration of the Sequence Alignment to Predict Across Species Susceptibility Tool for Rapid Assessment of Protein Conservation
Published on: February 10, 2023
Detecting the limits of regulatory element conservation and divergence estimation using pairwise and multiple
Daniel A Pollard1, Alan M Moses, Venky N Iyer
1Graduate Group in Biophysics, University of California, Berkeley, CA 94720, USA. dpollard@berkeley.edu
Multiple alignment accuracy in molecular evolution is mainly driven by the divergence between the two most distant species. This impacts inferences of transcription factor binding site conservation and divergence estimates, particularly on internal tree branches.
Area of Science:
- Molecular Evolution
- Bioinformatics
- Genomics
Background:
- Molecular evolutionary studies of noncoding sequences depend on multiple sequence alignments.
- The accuracy of these alignments and their impact on downstream inferences are not well understood across different conditions.
Purpose of the Study:
- To systematically examine multiple alignment accuracy and its effects on transcription factor binding site conservation and divergence estimation.
- To develop a simulation platform, CisEvolver, for modeling noncoding and transcription factor binding site evolution.
Main Methods:
- Developed CisEvolver, a simulation platform for molecular evolution.
- Used simulated alignments to assess multiple alignment accuracy.
- Evaluated the impact of alignment accuracy on transcription factor binding site conservation and divergence estimation.
Main Results:
- Multiple alignment accuracy is primarily determined by the divergence between the two most distantly related species.
- Conserved transcription factor binding sites can be misaligned even at short divergence distances, potentially confounding gain/loss studies.
- Divergence estimates are often overestimated at short distances and underestimated at long distances.
- Alignment accuracy varies significantly across phylogenetic tree branches, being least accurate for internal branches.
Conclusions:
- Variations in multiple alignment accuracy can introduce errors in molecular evolutionary inferences, mimicking biological variation.
- Findings inform species selection for analyses and highlight potential improvements for alignment and phylogenetic inference tools.
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