Dissection of Structural and Functional Requirements That Underlie the Interaction of ERdj3 Protein with Substrates in the Endoplasmic Reticulum

Otero, Joel; Lizák, Beáta; Feige, Matthias J.; Hendershot, Linda M.

doi:10.1074/jbc.m114.587147

Cited by 19 publications

(25 citation statements)

References 51 publications

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“…We defined the importance for domain II in ERdj3 tetramerization by monitoring the oligomeric state of ERdj3 mutants secreted from mammalian cells using gel filtration. We collected conditioned media from HEK293T cells transfected with ERdj3 mutants including the J‐domain mutant ERdj3 H53Q [which disrupts functional interactions between ERdj3 and BiP (Shen & Hendershot, )], ERdj3 ΔII (which completely removes the ERdj3 domain II), ERdj3 QEVV (a targeted deletion of residues 175–190 that constitute the two predicted β‐sheets shown in Fig A), ERdj3 ΔIII (a mutant lacking the entirety of domain III), and ERdj3 F326D [a mutant harboring a point mutation within domain III that disrupts ERdj3 oligomerization (Jin et al , ; Otero et al , )] (Figs B and EV2A). We also overexpressed two ERdj3 point mutants, ERdj3 T177P and ERdj3 Q186P , containing mutations predicted to disrupt the β‐sheets within domain II.…”

Section: Resultsmentioning

confidence: 99%

“…The EM map is deposited in the EM data bank (EMD-8707). Fig 3A), ERdj3 DIII (a mutant lacking the entirety of domain III), and ERdj3 F326D [a mutant harboring a point mutation within domain III that disrupts ERdj3 oligomerization (Jin et al, 2009;Otero et al, 2014)] (Figs 3B and EV2A). We also overexpressed two ERdj3 point mutants, ERdj3 T177P and ERdj3 Q186P , containing mutations predicted to disrupt the b-sheets within domain II.…”

Section: Targeted Deletion In Erdj3 Domain II Disrupts Formation Of Tmentioning

confidence: 99%

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The endoplasmic reticulum HSP40 co‐chaperone ERdj3/DNAJB11 assembles and functions as a tetramer

Chen

Chowdhury

et al. 2017

The EMBO Journal

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ERdj3/DNAJB11 is an endoplasmic reticulum (ER)-targeted HSP40 co-chaperone that performs multifaceted functions involved in coordinating ER and extracellular proteostasis. Here, we show that ERdj3 assembles into a native tetramer that is distinct from the dimeric structure observed for other HSP40 co-chaperones. An electron microscopy structural model of full-length ERdj3 shows that these tetramers are arranged as a dimer of dimers formed by distinct inter-subunit interactions involving ERdj3 domain II and domain III Targeted deletion of residues 175-190 within domain II renders ERdj3 a stable dimer that is folded and efficiently secreted from mammalian cells. This dimeric ERdj3 shows impaired substrate binding both in the ER and extracellular environments and reduced interactions with the ER HSP70 chaperone BiP. Furthermore, we show that overexpression of dimeric ERdj3 exacerbates ER stress-dependent reductions in the secretion of a destabilized, aggregation-prone protein and increases its accumulation as soluble oligomers in extracellular environments. These results reveal ERdj3 tetramerization as an important structural framework for ERdj3 functions involved in coordinating ER and extracellular proteostasis in the presence and absence of ER stress.

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Section: Resultsmentioning

confidence: 99%

Section: Targeted Deletion In Erdj3 Domain II Disrupts Formation Of Tmentioning

confidence: 99%

The endoplasmic reticulum HSP40 co‐chaperone ERdj3/DNAJB11 assembles and functions as a tetramer

Chen

Chowdhury

et al. 2017

The EMBO Journal

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“…Amongst the DnaJs involved in EC in this study, DNAJA2 is involved in refolding of disordered proteins (Baaklini et al, ), while others are mostly involved in ERAF. The ER‐associated chaperones (DNAJB12/ dnj‐1 , DNAJC3/ dnj‐7 , DNAJB11/ dnj‐20 , and SEC63/ dnj‐29 ) have been shown to have a function in ERAD (Grove et al, ; Oka et al, ; Otero, Lizák, Feige, & Hendershot, ; Ushioda et al, ; Figure ). Developmentally relevant proteins, such as signaling molecules and transcription factors, are known to be highly disordered (Ward, Sodhi, McGuffin, Buxton, & Jones, ), and such intrinsically disordered proteins are tightly regulated by degradation (Gsponer, Futschik, Teichmann, & Babu, ).…”

Section: Discussionmentioning

confidence: 99%

“…Amongst the DnaJs involved in EC in this study, DNAJA2 is involved in refolding of disordered proteins (Baaklini et al, 2012), while others are mostly involved in ERAF. The ER-associated chaperones (DNAJB12/dnj-1, DNAJC3/dnj-7, DNAJB11/dnj-20, and SEC63/dnj-29) have been shown to have a function in ERAD (Grove et al, 2011;Oka et al, 2013;Otero, Lizák, Feige, & Hendershot, 2014;Ushioda et al, 2008; Figure 8).…”

Section: Discussionmentioning

confidence: 99%

DnaJ chaperones contribute to canalization

et al. 2019

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Canalization, an intrinsic robustness of development to external (environmental) or internal (genetic) perturbations, was first proposed over half a century ago. However, whether the robustness to environmental stress (environmental canalization [EC]) and to genetic variation (genetic canalization) are underpinned by the same molecular basis remains elusive. The recent discovery of the involvement of two endoplasmic reticulum (ER)‐associated DnaJ genes in developmental buffering, orthologues of which are conserved across Metazoa, indicates that the role of ER‐associated DnaJ genes might be conserved across the animal kingdom. To test this, we surveyed the ER‐associated DnaJ chaperones in the nematode Caenorhabditis elegans. We then quantified the phenotype, in the form of variance and mean of seam cell counts, from RNA interference knockdown of DnaJs under three different temperatures. We find that seven out of eight ER‐associated DnaJs are involved in either EC or microenvironmental canalization. Moreover, we also found two DnaJ genes not specifically associated with ER (DNAJC2/dnj‐11 and DNAJA2/dnj‐19) were involved in canalization. Protein expression pattern showed that these DnaJs are upregulated by heat stress, yet not all of them are expressed in the seam cells. Moreover, we found that most of the buffering DnaJs also control lifespan. We therefore concluded that a number of DnaJ chaperones, not limited to those associated with the ER, are involved in canalization as a part of the complex system that underlies development.

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“…It is ubiquitously expressed, with the highest level of expression in secretory tissues, such as in the liver [Shen and Hendershot, 2005]. ERdj3 directly binds to nascent unfolded and misfolded proteins and transfers them to Hsp70 proteins, such as BiP [Guo and Snapp, 2013;Otero et al, 2014]. We demonstrate that depletion of ERdj3 reduces the accumulated ZAAT in the ER of the hepatocytes by enhancing ZAAT degradation.…”

mentioning

confidence: 82%

Erdj3 Has an Essential Role for Z Variant Alpha‐1‐Antitrypsin Degradation

Khodayari

Marek

et al. 2017

J of Cellular Biochemistry

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Alpha‐1‐antitrypsin deficiency (AATD) is an inherited disease characterized by emphysema and liver disease. AATD is most often caused by a single amino acid substitution at amino acid 342 in the mature protein, resulting in the Z mutation of the alpha‐1‐antitrypsin gene (ZAAT). This substitution is associated with misfolding and accumulation of ZAAT in the endoplasmic reticulum (ER) of hepatocytes and monocytes, causing a toxic gain of function. Retained ZAAT is eliminated by ER‐associated degradation and autophagy. We hypothesized that alpha‐1‐antitrypsin (AAT)‐interacting proteins play critical roles in quality control of human AAT. Using co‐immunoprecipitation, we identified ERdj3, an ER‐resident Hsp40 family member, as a part of the AAT trafficking network. Depleting ERdj3 increased the rate of ZAAT degradation in hepatocytes by redirecting ZAAT to the ER calreticulin‐EDEM1 pathway, followed by autophagosome formation. In the Huh7.5 cell line, ZAAT ER clearance resulted from enhancing ERdj3‐mediated ZAAT degradation by silencing ERdj3 while simultaneously enhancing autophagy. In this context, ERdj3 suppression may eliminate the toxic gain of function associated with polymerization of ZAAT, thus providing a potential new therapeutic approach to the treatment of AATD‐related liver disease. J. Cell. Biochem. 118: 3090–3101, 2017. © 2017 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals Inc.

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Dissection of Structural and Functional Requirements That Underlie the Interaction of ERdj3 Protein with Substrates in the Endoplasmic Reticulum

Cited by 19 publications

References 51 publications

The endoplasmic reticulum HSP40 co‐chaperone ERdj3/DNAJB11 assembles and functions as a tetramer

The endoplasmic reticulum HSP40 co‐chaperone ERdj3/DNAJB11 assembles and functions as a tetramer

DnaJ chaperones contribute to canalization

Erdj3 Has an Essential Role for Z Variant Alpha‐1‐Antitrypsin Degradation

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