ERdj3 regulates BiP occupancy in living cells

Feng Guo, Erik L. Snapp

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Co-chaperones regulate chaperone activities and are likely to impact a protein-folding environment as much as the chaperone itself. As co-chaperones are expressed substoichiometrically, the ability of co-chaperones to encounter a chaperone is crucial for chaperone activity. ERdj3, an abundant soluble endoplasmic reticulum (ER) co-chaperone of the Hsp70 BiP, stimulates the ATPase activity of BiP to increase BiP's affinity for client (or substrate) proteins. We investigated ERdj3 availability, how ERdj3 levels impact BiP availability, and the significance of J proteins for regulating BiP binding of clients in living cells. FRAP analysis revealed that overexpressed ERdj3-sfGFP dramatically decreases BiP-GFP mobility in a client-dependent manner. By contrast, ERdj3-GFP mobility remains low regardless of client protein levels. Native gels and co-immunoprecipitations established that ERdj3 associates with a large complex including Sec61α. Translocon binding probably ensures rapid encounters between emerging nascent peptides and stimulates BiP activity in the crucial early stages of secretory protein folding. Importantly, mutant BiP exhibited significantly increased mobility when it could not interact with any ERdjs. Thus, ERdjs appear to play the dual roles of increasing BiP affinity for clients and regulating delivery of clients to BiP. Our data suggest that BiP engagement of clients is enhanced in ER subdomains enriched in ERdj proteins.

Original languageEnglish (US)
Pages (from-to)1429-1439
Number of pages11
JournalJournal of Cell Science
Volume126
Issue number6
DOIs
StatePublished - Mar 15 2013

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Protein Folding
Endoplasmic Reticulum
Proteins
Immunoprecipitation
Adenosine Triphosphatases
Gels
Peptides

Keywords

  • Bip
  • Erdj
  • Fluorescence recovery after photobleaching
  • Frap
  • Quality control
  • Superfolder GFP

ASJC Scopus subject areas

  • Cell Biology

Cite this

ERdj3 regulates BiP occupancy in living cells. / Guo, Feng; Snapp, Erik L.

In: Journal of Cell Science, Vol. 126, No. 6, 15.03.2013, p. 1429-1439.

Research output: Contribution to journalArticle

Guo, Feng ; Snapp, Erik L. / ERdj3 regulates BiP occupancy in living cells. In: Journal of Cell Science. 2013 ; Vol. 126, No. 6. pp. 1429-1439.
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