A Cell Junctional Protein Network Associated with Connexin-26

International Journal of Molecular Sciences
Ana C BatissocoLuciana A Haddad

Abstract

GJB2 mutations are the leading cause of non-syndromic inherited hearing loss. GJB2 encodes connexin-26 (CX26), which is a connexin (CX) family protein expressed in cochlea, skin, liver, and brain, displaying short cytoplasmic N-termini and C-termini. We searched for CX26 C-terminus binding partners by affinity capture and identified 12 unique proteins associated with cell junctions or cytoskeleton (CGN, DAAM1, FLNB, GAPDH, HOMER2, MAP7, MAPRE2 (EB2), JUP, PTK2B, RAI14, TJP1, and VCL) by using mass spectrometry. We show that, similar to other CX family members, CX26 co-fractionates with TJP1, VCL, and EB2 (EB1 paralogue) as well as the membrane-associated protein ASS1. The adaptor protein CGN (cingulin) co-immuno-precipitates with CX26, ASS1, and TJP1. In addition, CGN co-immunoprecipitation with CX30, CX31, and CX43 indicates that CX association is independent on the CX C-terminus length or sequence. CX26, CGN, FLNB, and DAMM1 were shown to distribute to the organ of Corti and hepatocyte plasma membrane. In the mouse liver, CX26 and TJP1 co-localized at the plasma membrane. In conclusion, CX26 associates with components of other membrane junctions that integrate with the cytoskeleton.

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Citations

Aug 19, 2020·International Journal of Dermatology·Mun W LamPick N Woo
May 7, 2020·Frontiers in Neuroanatomy·E Anne MartinAdam C Miller
Feb 2, 2021·Frontiers in Nutrition·Ying DengHarry J Wichers
Jan 21, 2021·Biology·Laura Garcia-VegaPatricia E Martin
May 30, 2021·Proceedings of the National Academy of Sciences of the United States of America·Minwoo Wendy JangByung Yoon Choi

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Methods Mentioned

BETA
affinity capture
two-hybrid
nuclear translocation
affinity capture assay
immunoprecipitation
co-immunoprecipitation
PCR
electrophoresis

Software Mentioned

BioGRID
PSORT Homepage
Proteome
Scaffold
Clustal Omega
STRING
Hereditary Homepage Hydropathy
ZEN
Mouse Informatics
Mascot

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