ER-localized Shr3 is a selective co-translational folding chaperone necessary for amino acid permease biogenesis.
Proteins with multiple membrane-spanning segments (MS) co-translationally insert into the endoplasmic reticulum (ER) membrane of eukaryotic cells. Shr3, an ER membrane-localized chaperone in Saccharomyces cerevisiae, is required for the functional expression of a family of 18 amino acid permeases (AAP) comprised of 12 MS. We have used comprehensive scanning mutagenesis ... and deletion analysis of Shr3 combined with a modified split-ubiquitin approach to probe chaperone-substrate interactions in vivo. Shr3 selectively interacts with nested C-terminal AAP truncations in marked contrast to similar truncations of non-Shr3 substrate sugar transporters. Shr3-AAP interactions initiate with the first four MS of AAP and successively strengthen but weaken abruptly when all 12 MS are present. Shr3-AAP interactions are based on structural rather than sequence-specific interactions involving membrane and luminal domains of Shr3. The data align with Shr3 engaging nascent N-terminal chains of AAP, functioning as a scaffold to facilitate folding as translation completes.
Mesh Terms:
Amino Acid Transport Systems, Biological Transport, Endoplasmic Reticulum, Molecular Chaperones, Protein Folding, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Vesicular Transport Proteins
Amino Acid Transport Systems, Biological Transport, Endoplasmic Reticulum, Molecular Chaperones, Protein Folding, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Vesicular Transport Proteins
J Cell Biol
Date: Sep. 04, 2023
PubMed ID: 37477900
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