Usher syndrome proteins ADGRV1 (USH2C) and CIB2 (USH1J) interact and share a common interactome containing TRiC/CCT-BBS chaperonins

Abstract

The human Usher syndrome (USH) is the most common form of a sensory hereditary ciliopathy characterized by progressive vision and hearing loss. Mutations in the genes ADGRV1 and CIB2 have been associated with two distinct sub-types of USH, namely, USH2C and USH1J. The proteins encoded by the two genes belong to very distinct protein families: the adhesion G protein-coupled receptor ADGRV1 also known as the very large G protein-coupled receptor 1 (VLGR1) and the Ca2+- and integrin-binding protein 2 (CIB2), respectively. In the absence of tangible knowledge of the molecular function of ADGRV1 and CIB2, pathomechanisms underlying USH2C and USH1J are still unknown. Here, we aimed to enlighten the cellular functions of CIB2 and ADGRV1 by the identification of interacting proteins, a knowledge that is commonly indicative of cellular functions. Applying affinity proteomics by tandem affinity purification in combination with mass spectrometry, we identified novel potential binding partners of the CIB2 protein and compared these with the data set we previously obtained for ADGRV1. Surprisingly, the interactomes of both USH proteins showed a high degree of overlap indicating their integration in common networks, cellular pathways and functional modules which we confirmed by GO term analysis. Validation of protein interactions revealed that ADGRV1 and CIB2 mutually interact. In addition, we showed that the USH proteins also interact with the TRiC/CCT chaperonin complex and the Bardet Biedl syndrome (BBS) chaperonin-like proteins. Immunohistochemistry on retinal sections demonstrated the co-localization of the interacting partners at the photoreceptor cilia, supporting the role of USH proteins ADGRV1 and CIB2 in primary cilia function. The interconnection of protein networks involved in the pathogenesis of both syndromic retinal dystrophies BBS and USH suggest shared pathomechanisms for both syndromes on the molecular level.

Autoren

Joshua Linnert
Barbara Knapp
Baran E Güler
Karsten Boldt
Marius Ueffing
Uwe Wolfrum

DOI

10.3389/fcell.2023.1199069

eISSN

2296-634X

Zeitschrift

Frontiers in Cell and Developmental Biology

Online publication date

2023

Status

Published online

Herausgeber

Frontiers Media SA

Herausgeber URL

http://dx.doi.org/10.3389/fcell.2023.1199069

Datum der Datenerfassung

2023

Titel

Usher syndrome proteins ADGRV1 (USH2C) and CIB2 (USH1J) interact and share a common interactome containing TRiC/CCT-BBS chaperonins

Ausgabe der Zeitschrift

11

Data source: Crossref

Abstract

The human Usher syndrome (USH) is the most common form of a sensory hereditary ciliopathy characterized by progressive vision and hearing loss. Mutations in the genes ADGRV1 and CIB2 have been associated with two distinct sub-types of USH, namely, USH2C and USH1J. The proteins encoded by the two genes belong to very distinct protein families: the adhesion G protein-coupled receptor ADGRV1 also known as the very large G protein-coupled receptor 1 (VLGR1) and the Ca2+- and integrin-binding protein 2 (CIB2), respectively. In the absence of tangible knowledge of the molecular function of ADGRV1 and CIB2, pathomechanisms underlying USH2C and USH1J are still unknown. Here, we aimed to enlighten the cellular functions of CIB2 and ADGRV1 by the identification of interacting proteins, a knowledge that is commonly indicative of cellular functions. Applying affinity proteomics by tandem affinity purification in combination with mass spectrometry, we identified novel potential binding partners of the CIB2 protein and compared these with the data set we previously obtained for ADGRV1. Surprisingly, the interactomes of both USH proteins showed a high degree of overlap indicating their integration in common networks, cellular pathways and functional modules which we confirmed by GO term analysis. Validation of protein interactions revealed that ADGRV1 and CIB2 mutually interact. In addition, we showed that the USH proteins also interact with the TRiC/CCT chaperonin complex and the Bardet Biedl syndrome (BBS) chaperonin-like proteins. Immunohistochemistry on retinal sections demonstrated the co-localization of the interacting partners at the photoreceptor cilia, supporting the role of USH proteins ADGRV1 and CIB2 in primary cilia function. The interconnection of protein networks involved in the pathogenesis of both syndromic retinal dystrophies BBS and USH suggest shared pathomechanisms for both syndromes on the molecular level.

Addresses

Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University Mainz, Mainz, Germany.

Autoren

Joshua Linnert
Barbara Knapp
Baran E Güler
Karsten Boldt
Marius Ueffing
Uwe Wolfrum

DOI

10.3389/fcell.2023.1199069

eISSN

2296-634X

Externe Identifier

PubMed Identifier: 37427378
PubMed Central ID: PMC10323441

Funding acknowledgements

Johannes Gutenberg-Universität Mainz:
Foundation Fighting Blindness:
Deutsche Forschungsgemeinschaft:

Open access

true

ISSN

2296-634X

Zeitschrift

Frontiers in cell and developmental biology

Sprache

eng

Medium

Electronic-eCollection

Online publication date

2023

Open access status

Open Access

Paginierung

1199069

Datum der Veröffentlichung

2023

Status

Published

Publisher licence

CC BY

Datum der Datenerfassung

2023

Titel

Usher syndrome proteins ADGRV1 (USH2C) and CIB2 (USH1J) interact and share a common interactome containing TRiC/CCT-BBS chaperonins.

Sub types

research-article
Journal Article

Ausgabe der Zeitschrift

11

Files

https://europepmc.org/articles/PMC10323441?pdf=render

Data source: Europe PubMed Central

Abstract

The human Usher syndrome (USH) is the most common form of a sensory hereditary ciliopathy characterized by progressive vision and hearing loss. Mutations in the genes ADGRV1 and CIB2 have been associated with two distinct sub-types of USH, namely, USH2C and USH1J. The proteins encoded by the two genes belong to very distinct protein families: the adhesion G protein-coupled receptor ADGRV1 also known as the very large G protein-coupled receptor 1 (VLGR1) and the Ca2+- and integrin-binding protein 2 (CIB2), respectively. In the absence of tangible knowledge of the molecular function of ADGRV1 and CIB2, pathomechanisms underlying USH2C and USH1J are still unknown. Here, we aimed to enlighten the cellular functions of CIB2 and ADGRV1 by the identification of interacting proteins, a knowledge that is commonly indicative of cellular functions. Applying affinity proteomics by tandem affinity purification in combination with mass spectrometry, we identified novel potential binding partners of the CIB2 protein and compared these with the data set we previously obtained for ADGRV1. Surprisingly, the interactomes of both USH proteins showed a high degree of overlap indicating their integration in common networks, cellular pathways and functional modules which we confirmed by GO term analysis. Validation of protein interactions revealed that ADGRV1 and CIB2 mutually interact. In addition, we showed that the USH proteins also interact with the TRiC/CCT chaperonin complex and the Bardet Biedl syndrome (BBS) chaperonin-like proteins. Immunohistochemistry on retinal sections demonstrated the co-localization of the interacting partners at the photoreceptor cilia, supporting the role of USH proteins ADGRV1 and CIB2 in primary cilia function. The interconnection of protein networks involved in the pathogenesis of both syndromic retinal dystrophies BBS and USH suggest shared pathomechanisms for both syndromes on the molecular level.

Date of acceptance

2023

Autoren

Joshua Linnert
Barbara Knapp
Baran E Güler
Karsten Boldt
Marius Ueffing
Uwe Wolfrum

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/37427378

DOI

10.3389/fcell.2023.1199069

Externe Identifier

PubMed Central ID: PMC10323441

ISSN

2296-634X

Zeitschrift

Front Cell Dev Biol

Schlüsselwörter

TRiC/CCT chaperonins
VLGR1
bardet biedl syndrome (BBS)
photoreceptor cells
primary cilia
protein networks
retinal ciliopathies
usher syndrome

Sprache

eng

Country

Switzerland

Paginierung

1199069

PII

1199069

Datum der Veröffentlichung

2023

Status

Published online

Titel

Usher syndrome proteins ADGRV1 (USH2C) and CIB2 (USH1J) interact and share a common interactome containing TRiC/CCT-BBS chaperonins.

Sub types

Journal Article

Ausgabe der Zeitschrift

11

Data source: PubMed

Author's licence

CC-BY

Autoren

Joshua Linnert
Barbara Knapp
Baran E Güler
Karsten Boldt
Marius Ueffing
Uwe Wolfrum

Hosting institution

Universitätsbibliothek Mainz

Sammlungen

DFG-491381577-G

Resource version

Published version

DOI

10.3389/fcell.2023.1199069

File(s) embargoed

false

Open access

true

ISSN

2296-634X

Zeitschrift

Frontiers in Cell and Developmental Biology

Schlüsselwörter

570 Biowissenschaften
570 Life sciences

Sprache

eng

Open access status

Open Access

Paginierung

1199069

Datum der Veröffentlichung

2023

Public URL

https://openscience.ub.uni-mainz.de/handle/20.500.12030/9276

Herausgeber

Frontiers

Datum der Datenerfassung

2023

Datum, an dem der Datensatz öffentlich gemacht wurde

2023

Zugang

Public

Titel

Usher syndrome proteins ADGRV1 (USH2C) and CIB2 (USH1J) interact and share a common interactome containing TRiC/CCT-BBS chaperonins

Ausgabe der Zeitschrift

11

Files

usher_syndrome_proteins_adgrv-20230706092555440.pdf

Data source: OPENSCIENCE.UB

Usher syndrome proteins ADGRV1 (USH2C) and CIB2 (USH1J) interact and share a common interactome containing TRiC/CCT-BBS chaperonins

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