title: Members of the Hsp70 Family Recognize Distinct Types of Sequences to Execute ER Quality Control.
abstract: Protein maturation in the endoplasmic reticulum is controlled by multiple chaperones, but how they recognize and determine the fate of their clients remains unclear. We developed an in vivo peptide library covering substrates of the ER Hsp70 system: BiP, Grp170, and three of BiP's DnaJ-family co-factors (ERdj3, ERdj4, and ERdj5). In vivo binding studies revealed that sites for pro-folding chaperones BiP and ERdj3 were frequent and dispersed throughout the clients, whereas Grp170, ERdj4, and ERdj5 specifically recognized a distinct type of rarer sequence with a high predicted aggregation potential. Mutational analyses provided insights into sequence recognition characteristics for these pro-degradation chaperones, which could be readily introduced or disrupted, allowing the consequences for client fates to be determined. Our data reveal unanticipated diversity in recognition sequences for chaperones; establish a sequence-encoded interplay between protein folding, aggregation, and degradation; and highlight the ability of clients to co-evolve with chaperones, ensuring quality control.
authors.affiliation: Department of Tumor Cell Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USA.
authors.forename: Julia
authors.initials: J
authors.lastname: Behnke
authors.affiliation: Department of Tumor Cell Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USA.
authors.forename: Melissa J
authors.initials: MJ
authors.lastname: Mann
authors.affiliation: Department of Tumor Cell Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USA; Department of Chemistry, Rhodes College, 2000 N. Parkway, Memphis, TN 38112, USA.
authors.forename: Fei-Lin
authors.initials: FL
authors.lastname: Scruggs
authors.affiliation: Department of Tumor Cell Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USA; Center for Integrated Protein Science at the Department of Chemistry and Institute for Advanced Study, Technische Universität München, Lichtenbergstr. 4, 85748 Garching, Germany. Electronic address: matthias.feige@tum.de.
authors.forename: Matthias J
authors.initials: MJ
authors.lastname: Feige
authors.affiliation: Department of Tumor Cell Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USA. Electronic address: linda.hendershot@stjude.org.
authors.forename: Linda M
authors.initials: LM
authors.lastname: Hendershot
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002522:Chlorocebus aethiops; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D000091342:Endoplasmic Reticulum Chaperone BiP; D015870:Gene Expression; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D050956:HSP40 Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018832:Molecular Chaperones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D017510:Protein Folding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D014162:Transfection; D019076:Transgenes
chemical_list: C494648:DNAJB11 protein, human; C478021:DNAJB9 protein, human; C471179:DNAJC10 protein, human; D000091342:Endoplasmic Reticulum Chaperone BiP; D006023:Glycoproteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D008565:Membrane Proteins; D018832:Molecular Chaperones; D019151:Peptide Library; C098410:glucose-regulated protein 170