Transmembrane proteins have unique requirements to fold and integrate into the endoplasmic reticulum (ER) membrane. Most notably, transmembrane proteins must fold in three separate environments: extracellular domains fold in the oxidizing environment of the ER lumen, transmembrane domains (TMDs) fold within the lipid bilayer, and cytosolic domains fold in the reducing environment of the cytosol. Moreover, each region is acted upon by a unique set of chaperones and monitored by components of the ER associated quality control machinery that identify misfolded domains in each compartment. One factor is the ER lumenal Hsp70-like chaperone, Lhs1. Our previous work established that Lhs1 is required for the degradation of the unassembled α-subunit of the epithelial sodium channel (αENaC), but not the homologous β- and γENaC subunits. However, assembly of the ENaC heterotrimer blocked the Lhs1-dependent ER associated degradation (ERAD) of the α-subunit, yet the characteristics that dictate the specificity of Lhs1-dependent ERAD substrates remained unclear. We now report that Lhs1-dependent substrates share a unique set of features. First, all Lhs1 substrates appear to be unglycosylated, and second they contain two TMDs. Each substrate also contains orphaned or unassembled TMDs. Additionally, interfering with inter-subunit assembly of the ENaC trimer results in Lhs1-dependent degradation of the entire complex. Finally, our work suggests that Lhs1 is required for a subset of ERAD substrates that also require the Hrd1 ubiquitin ligase. Together, these data provide hints as to the identities of as-yet unconfirmed substrates of Lhs1 and potentially of the Lhs1 homolog in mammals, GRP170.
Lhs1 dependent ERAD is determined by transmembrane domain context
Maria Sukhoplyasova, Abigail M. Keith, Emma M. Perrault, Hannah E. Vorndran, Alexa S. Jordahl, Megan E. Yates, Ashutosh Pastor, Zachary Li, Michael L. Freaney, Riddhi A. Deshpande, David B. Adams, Christopher J. Guerriero, Shujie Shi, Thomas R. Kleyman, Ossama B. Kashlan, Jeffrey L. Brodsky, Teresa M. Buck; Lhs1 dependent ERAD is determined by transmembrane domain context. Biochem J 27 September 2023; 480 (18): 1459–1473. doi: https://doi.org/10.1042/BCJ20230075
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