Competitive Inhibition of the Endoplasmic Reticulum Signal Peptidase by Non-cleavable Mutant Preprotein Cargos

Cui, Jingqiu; Chen, Wei; Sun, Jin; Guo, Huan; Madley, Rachel; Xiong, Yi; Pan, X. Q.; Wang, Hongliang; Tai, Andrew W.; Weiss, Michael A.; Arvan, Peter; Liu, Ming

doi:10.1074/jbc.m115.692350

Cited by 26 publications

(20 citation statements)

References 47 publications

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“…We observed shared interactors between OC43 and SARS-CoV-2, such as the Nglycosylation factor RFT1 39,40 and a sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA -ATP2A2) 41 . In addition, we identified shared interactors between OC43 and SARS-CoV-1, including a regulator of UPR-mediated apoptosis (WLS or GPR177) 42 , a member of the signal peptidase complex (SEC11A) 43 , and factors involved in cholesterol synthesis (IDI1, DHCR7) [44][45][46][47] . WLS, SEC11A, and DHRC7 exhibited higher enrichment for OC43, whereas IDI1 was more greatly enriched for SARS-CoV-1.…”

Section: Analysis Of Go-mentioning

confidence: 99%

Comparative multiplexed interactomics of SARS-CoV-2 and homologous coronavirus non-structural proteins identifies unique and shared host-cell dependencies

Davies

Almasy

McDonald

et al. 2020

Preprint

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Human coronaviruses (hCoV) have become a threat to global health and society, as evident from the SARS outbreak in 2002 caused by SARS-CoV-1 and the most recent COVID-19 pandemic caused by SARS-CoV-2. Despite high sequence similarity between SARS-CoV-1 and -2, each strain has distinctive virulence. A better understanding of the basic molecular mechanisms mediating changes in virulence is needed. Here, we profile the virus-host protein-protein interactions of two hCoV non-structural proteins (nsps) that are critical for virus replication. We use tandem mass tag-multiplexed quantitative proteomics to sensitively compare and contrast the interactomes of nsp2 and nsp4 from three betacoronavirus strains: SARS-CoV-1, SARS-CoV-2, and hCoV-OC43 - an endemic strain associated with the common cold. This approach enables the identification of both unique and shared host cell protein binding partners and the ability to further compare the enrichment of common interactions across homologs from related strains. We identify common nsp2 interactors involved in endoplasmic reticulum (ER) Ca2+ signaling and mitochondria biogenesis. We also identifiy nsp4 interactors unique to each strain, such as E3 ubiquitin ligase complexes for SARS-CoV-1 and ER homeostasis factors for SARS-CoV-2. Common nsp4 interactors include N-linked glycosylation machinery, unfolded protein response (UPR) associated proteins, and anti-viral innate immune signaling factors. Both nsp2 and nsp4 interactors are strongly enriched in proteins localized at mitochondrial-associated ER membranes suggesting a new functional role for modulating host processes, such as calcium homeostasis, at these organelle contact sites. Our results shed light on the role these hCoV proteins play in the infection cycle, as well as host factors that may mediate the divergent pathogenesis of OC43 from SARS strains. Our mass spectrometry workflow enables rapid and robust comparisons of multiple bait proteins, which can be applied to additional viral proteins. Furthermore, the identified common interactions may present new targets for exploration by host-directed anti-viral therapeutics.

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Section: Analysis Of Go-mentioning

confidence: 99%

Comparative multiplexed interactomics of SARS-CoV-2 and homologous coronavirus non-structural proteins identifies unique and shared host-cell dependencies

Davies

Almasy

McDonald

et al. 2020

Preprint

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“…This would suggest that the residues downstream of the cleavage site play no role for cleavage efficiency. However, a proline next to the cleavage site does inhibit cleavage and residues further downstream can influence signal‐peptide cleavage; for example, HIV‐1 Env , HCMV US11 and EDEM1 lose their signal peptides post‐translationally, effectively acting as type II signal anchors for the early period of the protein's life. Cleavage of HIV‐1 Env required some folding , suggesting interplay between the signal peptide and the folding molecule, while delayed cleavage of EDEM1 affects its substrate specificity.…”

Section: Making An Entry – the Role Of Signal Peptide Cleavagementioning

confidence: 99%

Co‐ and Post‐Translational Protein Folding in the ER

Ellgaard

McCaul

Chatsisvili

et al. 2016

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The biophysical rules that govern folding of small, single-domain proteins in dilute solutions are now quite well understood. The mechanisms underlying co-translational folding of multidomain and membrane-spanning proteins in complex cellular environments are often less clear. The endoplasmic reticulum (ER) produces a plethora of membrane and secretory proteins, which must fold and assemble correctly before ER exit -if these processes fail, misfolded species accumulate in the ER or are degraded. The ER differs from other cellular organelles in terms of the physicochemical environment and the variety of ER-specific protein modifications. Here, we review chaperone-assisted co-and post-translational folding and assembly in the ER and underline the influence of protein modifications on these processes. We emphasize how method development has helped advance the field by allowing researchers to monitor the progression of folding as it occurs inside living cells, while at the same time probing the intricate relationship between protein modifications during folding.

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“…translocation is coupled to protein synthesis) using a process in which the ribonucleoprotein signal recognition particle (7-9) can interact with the SP (10,11) and initiate delivery of the elongating polypeptide chain to the Sec61 translocon. As the nascent polypeptides enter the ER lumen, the SPs are efficiently removed by signal peptidase on the lumenal side of the ER membrane (12)(13)(14).…”

mentioning

confidence: 99%

Positive charge in the n-region of the signal peptide contributes to efficient post-translational translocation of small secretory preproteins

Guo

Sun

et al. 2018

Journal of Biological Chemistry

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Increasing evidence indicates that many small secretory preproteins can undergo post-translational translocation across the membrane of the endoplasmic reticulum. Although the cellular machinery involved in post-translational translocation of small secretory preproteins has begun to be elucidated, the intrinsic signals contained within these small secretory preproteins that contribute to their efficient post-translational translocation remain unknown. Here, we analyzed the eukaryotic secretory proteome and discovered the small secretory preproteins tend to have a higher probability to harbor the positive charge in the n-region of the signal peptide (SP). Eliminating the positive charge of the n-region blocked post-translational translocation of newly synthesized preproteins and selectively impaired translocation efficiency of small secretory preproteins. The pathophysiological significance of the positive charge in the n-region of SP was underscored by recently identified preproinsulin SP mutations that impair translocation of preproinsulin and cause maturity onset diabetes of youth (MODY). Remarkably, we have found that slowing the polypeptide elongation rate of small secretory preproteins could alleviate the translocation defect caused by loss of the n-region positive charge of the signal peptide. Together, these data reveal not only a previously unrecognized role of the n-region's positive charge in ensuring efficient post-translational translocation of small secretory preproteins, but they also highlight the molecular contribution of defects in this process to the pathogenesis of genetic disorders such as MODY.

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Competitive Inhibition of the Endoplasmic Reticulum Signal Peptidase by Non-cleavable Mutant Preprotein Cargos

Cited by 26 publications

References 47 publications

Comparative multiplexed interactomics of SARS-CoV-2 and homologous coronavirus non-structural proteins identifies unique and shared host-cell dependencies

Comparative multiplexed interactomics of SARS-CoV-2 and homologous coronavirus non-structural proteins identifies unique and shared host-cell dependencies

Co‐ and Post‐Translational Protein Folding in the ER

Positive charge in the n-region of the signal peptide contributes to efficient post-translational translocation of small secretory preproteins

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