Standard, Stand-alone, & Ecosystem Analyses#

HyPhy is designed as a modular, extensible evolutionary analysis engine. While selection analyses are its most famous application, the platform supports a wide array of built-in CLI standard analyses, community batch files (hyphy-analyses), and direct integration with web platforms like Galaxy.


1. Built-in CLI Standard Analyses#

When running HyPhy from the command line, you can execute a wide variety of standard analyses directly. Running hyphy -i launches an interactive prompt, or you can run any analysis headlessly by passing its command name (viewable via hyphy --help).

Below is an overview of the core built-in analyses:

Command Analysis Name Description
busted BUSTED Test for episodic gene-wide selection.
busted-ph BUSTED-Phenotype Test if episodic selection is associated with phenotype/foreground branches.
absrel aBSREL Test for lineage-specific episodic positive selection using an adaptive branch-site model.
fel FEL Identify individual sites subject to pervasive selection using a maximum likelihood approach.
fubar FUBAR High-power Bayesian method for detecting site-level pervasive selection on large alignments.
meme MEME Mixed Effects Model of Evolution to find site-level episodic and pervasive positive selection.
slac SLAC Counting-based site-level selection test using joint maximum likelihood ancestral reconstruction.
relax RELAX Test for selection relaxation or intensification along a designated set of branches.
prime PRIME Property-Informed Models of Evolution to test if selection preserves or alters amino acid properties.
gard GARD Heuristic search for recombination breakpoints and partitioned tree reconstruction.
contrast-fel Contrast-FEL Test for site-level differences in selective pressures between predefined clades.
contrast-meme Contrast-MEME Mixed-effects test for site-level selective differences between clades.
bgm BGM Apply Bayesian Graphical Models to detect coevolutionary interactions between codon positions.
fmm FitMultiModel Fit models permitting double or triple instantaneous nucleotide substitutions.
fst F_ST Compute population genetic differentiation measures with permutation testing.
sm Slatkin-Maddison Test for gene flow and estimate migration events across populations.
leisr LEISR Infer relative evolutionary rates on nucleotide/protein alignments (similar to Rate4Site).
sw / spl Sliding Window / SimPlot Map genetic similarity/distance along alignments using a sliding window.
cln Clean Names Sanitize sequence headers and replace stop codons with gaps.
conv Convert Codons Translate an in-frame codon multiple sequence alignment to proteins.
label-tree Label Branches Annotate branches in a Newick tree for downstream partition analyses.

2. Galaxy Ecosystem Integration#

The Galaxy Project is a web-based platform for accessible, reproducible, and transparent computational biomedical research. Galaxy is a primary partner in the HyPhy ecosystem, providing a user-friendly graphical interface for high-performance clusters without requiring command-line usage.

The Galaxy Intergalactic Utilities Commission (IUC) maintains official, robust wrapper XMLs for HyPhy tools under the tools-iuc repository. The local equivalents are stored in galaxy-tools/tools-iuc/tools/hyphy.

Supported Galaxy XML Wrappers:#

  • Selection Tests: hyphy_busted.xml, hyphy_absrel.xml, hyphy_fel.xml, hyphy_fubar.xml, hyphy_meme.xml, hyphy_slac.xml
  • Clade-Specific & Property Tests: hyphy_cfel.xml (Contrast-FEL), hyphy_prime.xml (PRIME)
  • Recombination & Relaxation: hyphy_gard.xml, hyphy_relax.xml
  • Utility & Networks: hyphy_bgm.xml (Coevolution), hyphy_cln.xml (Data prep), hyphy_conv.xml (Translation)

By packaging parameter validations, automatic test suites (in /test-data), and detailed biological guidelines directly inside these wrappers, Galaxy enables seamless integration of HyPhy into multi-tool automated workflows.


3. Community Stand-alone Analyses (hyphy-analyses)#

For advanced pipelines, custom models, and experimental methods, the HyPhy team maintains the hyphy-analyses repository. These analyses can be run locally using the HyPhy command-line engine by passing the path to their corresponding batch file.

Selection & Modeling Extensions#

  • BUSTED-PH: Test if episodic diversifying selection is associated with a specific phenotype/trait on foreground branches.
  • BUSTED-MH: BUSTED variant supporting multiple instantaneous nucleotide substitutions.
  • BUSTED-SR: BUSTED variant incorporating synonymous rate variation.
  • RELAX-joint: Joint RELAX analysis across multiple gene alignments to boost statistical power.
  • RELAX-scan: Perform sliding-window scans of selection relaxation along a gene.
  • MulticlassSynonymousSubstitutions: Model codon evolution with multiple distinct classes of synonymous codons.

Model Fitting & Emulation#

  • FitMG94: Maximum likelihood fit of a standard Muse-Gaut 94 codon model.
  • FitMultiModel: Fit and compare models with single, double, and triple instantaneous mutations.
  • PAML-emulator: Emulate standard PAML models (such as M0, M1a, M2a, M7, M8) directly within the HyPhy framework.
  • NucleotideNonREV: Fit non-reversible nucleotide models of sequence evolution.

Simulation & Synthetic Data#

  • SimulateMG94: Simulate synthetic codon alignments along a tree under MG94 models.
  • SimulateProtein: Simulate amino acid sequence evolution under standard matrices.

Sequence Preparation & Utilities#

  • codon-msa: A helper workflow for generating codon-aware multiple sequence alignments.
  • LabelTrees: Automatically annotate phylogenetic trees for downstream branch-partition analyses.
  • clean-names / remove-duplicates: Sanitization scripts to clean sequence headers and prune identical taxa from trees and alignments.
  • extract-partitions: Extract multi-partition alignments into individual files.