# PharaohFUN: phylogenomic analysis for plant protein history and function elucidation

**Authors:** Marcos Ramos-González, Víctor Ramos-González, Emma Serrano-Pérez, Christina Arvanitidou, Jorge Hernández-García, Mercedes García-González, Francisco J Romero-Campero

PMC · DOI: 10.1093/molbev/msag011 · Molecular Biology and Evolution · 2026-01-31

## TL;DR

PharaohFUN is a web tool for analyzing plant protein evolution and function using phylogenomic data from diverse photosynthetic organisms.

## Contribution

PharaohFUN introduces a unified phylogenomic framework for plant proteins, integrating diverse species and functional analysis tools.

## Key findings

- PharaohFUN enables evolutionary analysis of plant proteins across land plants and microalgae.
- The tool successfully traced evolutionary histories of Arabidopsis transcription factors and CCA1 protein.
- PharaohFUN provides accessible phylogenomic analysis for users without bioinformatics training.

## Abstract

Since DNA sequencing has become commonplace, the development of efficient methods and tools to explore gene sequences has become indispensable. In particular, despite photosynthetic eukaryotes constituting the largest percentage of terrestrial biomass, computational functional characterization of gene sequences in these organisms still predominantly relies on comparisons with Arabidopsis thaliana and other angiosperms. This paper introduces PharaohFUN, a web application designed for the evolutionary and functional analysis of protein sequences in photosynthetic eukaryotes, leveraging orthology relationships between them. PharaohFUN incorporates a homogeneous representative sampling of key species in this group, bridging clades that have traditionally been studied separately, thus establishing a comprehensive evolutionary framework to draw conclusions about sequence evolution and function. For this purpose, it incorporates modules for exploring gene tree evolutionary history, expansion and contraction events, ancestral states, domain identification, multiple sequence alignments, and diverse functional annotation. It also incorporates different search modes to facilitate its use and increase its reach within the community. Tests were performed on the whole transcription factor toolbox of A. thaliana and on CCA1 protein to assess its utility for both large-scale and fine-grained phylogenetic studies. These exemplify how PharaohFUN accurately traces the corresponding evolutionary histories of these proteins by unifying results for land plants, streptophyte and chlorophyte microalgae. Thus, PharaohFUN democratices access to these kinds of analyses in photosynthetic organisms for every user, independently of their prior training in bioinformatics.

Graphical Abstract

## Linked entities

- **Genes:** CCA1 (cataract, congenital, cerulean type, 1) [NCBI Gene 878]
- **Proteins:** CCA1 (cataract, congenital, cerulean type, 1)
- **Species:** Arabidopsis thaliana (taxon 3702)

## Full-text entities

- **Genes:** CCA1 (circadian clock associated 1) [NCBI Gene 819296] {aka AtCCA1, F19D11.11, MYB-RELATED DNA BINDING PROTEIN, circadian clock associated 1}, ZAT6 (6) [NCBI Gene 830313] {aka AtZAT6, C2H2, C2H2 ZINC FINGER TRANSCRIPTION FACTOR, COLD INDUCED ZINC FINGER PROTEIN 2, CZF2, T19N18.70}, WRKY19 (protein kinase family protein) [NCBI Gene 826810] {aka ATWRKY19, F16J13.90, F16J13_90, MAPK/ERK KINASE KINASE 4, MAPKKK11, MEKK4}, BES1 (Brassinosteroid signaling positive regulator (BZR1) family protein) [NCBI Gene 838518] {aka 107 PROTEIN, BRASSINAZOLE-RESISTANT 2, BRI1-EMS-SUPPRESSOR 1, BZR2, F18O14.7, F18O14_7}, LHY (Homeodomain-like superfamily protein) [NCBI Gene 839341] {aka LATE ELONGATED HYPOCOTYL, LATE ELONGATED HYPOCOTYL 1, LHY1, T25K16.6, T25K16_6}, LSD1 (LSD1 zinc finger family protein) [NCBI Gene 827786] {aka F9F13.30, F9F13_30, LESION SIMULATING DISEASE, LESION SIMULATING DISEASE 1}, BZR1 (Brassinosteroid signaling positive regulator (BZR1) family protein) [NCBI Gene 843845] {aka BRASSINAZOLE-RESISTANT 1, F9E10.7, F9E10_7}, PHYB (phytochrome B) [NCBI Gene 816394] {aka HY3, MSF3.17, MSF3_17, OOP1, OUT OF PHASE 1, PHYTOCHROME B}, TOC1 (CCT motif -containing response regulator protein) [NCBI Gene 836259] {aka APRR1, AtTOC1, MFB13.13, MFB13_13, PRR1, PSEUDO-RESPONSE REGULATOR 1}, TIR (toll/interleukin-1 receptor-like protein) [NCBI Gene 843624] {aka AtTN10, F3N23.13, F3N23_13, TIR-nucleotide binding site family 10, TN10, toll/interleukin-1 receptor-like}
- **Chemicals:** auxin (MESH:D007210), OG (-)
- **Species:** Cymbomonas tetramitiformis (species) [taxon 36881], Brassica rapa (field mustard, species) [taxon 3711], Sorghum bicolor (broomcorn, species) [taxon 4558], Chlorophyta (green algae, phylum) [taxon 3041], Mesostigma viride (species) [taxon 41882], Ostreococcus tauri (species) [taxon 70448], Abies alba (abete bianco, species) [taxon 45372], Alsophila spinulosa (species) [taxon 204586], PX clade (clade) [taxon 569578], Chromochloris zofingiensis (species) [taxon 31302], Spirogloea muscicola (species) [taxon 2576442], Solanum lycopersicum (tomato, species) [taxon 4081], Marchantia polymorpha (common liverwort, species) [taxon 3197], Zygnema circumcarinatum (species) [taxon 35869], Physcomitrium patens (species) [taxon 3218], Klebsormidium nitens (species) [taxon 105231], Bathycoccus prasinos (species) [taxon 41875], Helianthus annuus (common sunflower, species) [taxon 4232], Rhodophyta (red algae, phylum) [taxon 2763], Amaranthus hypochondriacus (grain amaranth, species) [taxon 28502], Oryza sativa (Asian cultivated rice, species) [taxon 4530], Arabidopsis thaliana (mouse-ear cress, species) [taxon 3702], S. bicolor [taxon 381118], Volvox carteri (species) [taxon 3067], Mesotaenium kramstae (species) [taxon 184486]
- **Cell lines:** B. rapa — Opodiphthera eucalypti (Emperor gum moth), Spontaneously immortalized cell line (CVCL_C2VY)

## Full text

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## Figures

6 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12866927/full.md

## References

82 references — full list in the complete paper: https://tomesphere.com/paper/PMC12866927/full.md

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Source: https://tomesphere.com/paper/PMC12866927