Conserved Gene Microsynteny Unveils Functional Interaction Between Protein Disulfide Isomerase and Rho Guanine-Dissociation Inhibitor Families

Ana I S Moretti,Matthias S Leisegang,Carolina G Fernandes,Matheus H Dias,Ngoc-Vinh Tran,João Wosniak,Denise C Fernandes,Ralf P Brandes,Patrícia Nolasco,Jessyca C Pavanelli,Ingo Ebersberger,Francisco R M Laurindo,Leonardo Y Tanaka,Diego Bonatto,Daniela Kajihara,Hanjoong Jo

doi:10.1038/s41598-017-16947-5

Abstract

Protein disulfide isomerases (PDIs) support endoplasmic reticulum redox protein folding and cell-surface thiol-redox control of thrombosis and vascular remodeling. The family prototype PDIA1 regulates NADPH oxidase signaling and cytoskeleton organization, however the related underlying mechanisms are unclear. Here we show that genes encoding human PDIA1 and its two paralogs PDIA8 and PDIA2 are each flanked by genes encoding Rho guanine-dissociation inhibitors (GDI), known regulators of RhoGTPases/cytoskeleton. Evolutionary histories of these three microsyntenic regions reveal their emergence by two successive duplication events of a primordial gene pair in the last common vertebrate ancestor. The arrangement, however, is substantially older, detectable in echinoderms, nematodes, and cnidarians. Thus, PDI/RhoGDI pairing in the same transcription orientation emerged early in animal evolution and has been largely maintained. PDI/RhoGDI pairs are embedded into conserved genomic regions displaying common cis-regulatory elements. Analysis of gene expression datasets supports evidence for PDI/RhoGDI coexpression in developmental/inflammatory contexts. PDIA1/RhoGDIα were co-induced in endothelial cells upon CRISP-R-promoted transcription activation of each pair component, and also in mouse arterial intima during flow-induced remodeling. We provide evidence for physical interaction between both proteins. These data support strong functional links between PDI and RhoGDI families, which likely maintained PDI/RhoGDI microsynteny along > 800-million years of evolution.

Highlights

Such as cell adhesion, protease shedding, thrombosis, platelet activation[1,2,3,4,5,6,7], vascular remodeling and mechanoadaptation[8]
We have identified orthologs to 8 of these genes in a selection of taxa spanning the eukaryotic tree of life and reconstructed the evolutionary history of this gene family using maximum likelihood (Supplementary Fig. S1)
We subsequently reconstructed the evolutionary history of the RhoGDI gene family, and a picture strikingly similar to that of PDIA1 family emerged (Supplementary Figure 2)

Summary

Introduction

Such as cell adhesion, protease shedding, thrombosis, platelet activation[1,2,3,4,5,6,7], vascular remodeling and mechanoadaptation[8]. The nature of PDIA1 and RhoGDIα interaction is unknown, and their association is not obvious, given their distinct canonical subcellular locations and functions Further investigating their cooperation at several hierarchic levels is important to establish plausible mechanistic links between PDIA1 and cytoskeleton/RhoGTPase regulation. Further investigation revealed that the evolutionary maintenance of the PDI/RhoGDI gene microsynteny correlates with their functional cooperation, documented by evidences of coregulation, coexpression and physical interaction of their protein products. Such functional interactions may have constrained the disruption of their syntenic locations along evolution. These data further support strong mechanistic links between PDI and RhoGDI/RhoGTPase families potentially affecting cytoskeletal organization

Methods

Results

Conclusion