Expression of three topologically distinct membrane proteins elicits unique stress response pathways in the yeast<i>Saccharomyces cerevisiae</i>

Buck, Teresa M.; Jordan, Rick; Lyons‐Weiler, James; Adelman, Joshua L.; Needham, Patrick G.; Kleyman, Thomas R.; Brodsky, Jeffrey L.

doi:10.1152/physiolgenomics.00101.2014

Cited by 10 publications

(10 citation statements)

References 171 publications

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“…Compared to controls with D542-UPR alone, or treated with DTT to fully induce the UPR (42), both NC and OPT expression activated the UPR response, but the level of activation was significantly higher for OPT ( Figure 2D). UPR activation has been observed during expression of membrane protein mutants that destabilize the native structure or during over-production of specific membrane proteins for crystallographic purposes (43,44). Although Opt was not misfolded ( Figure 2B) and its concentration was not significantly higher than NC ( Figure 2C), activation of the UPR stress response suggests that post-transcriptional or translational events during expression of OPT may be responsible for the slow-growth phenotype…”

Section: Resultsmentioning

confidence: 99%

“…Our results suggest that RAC-Ssb may be responsible for this role in the context of at least MFS class of membrane proteins. When rapidly translated membrane proteins such as Opt are highly expressed, RAC-Ssb interactions with these nascent chains and the subsequent degradation of a subset of these polypeptides could overwhelm the basal level of protein quality control in cells, which would increase the proteasomal load and trigger cell stress and possibly the UPR (44,71). Notably, transporters were highly enriched in the subset of proteins identified to be toxic in a genome-wide analysis of protein overexpression in yeast (74), consistent with our model for RAC-Ssb function.…”

Section: Discussionmentioning

confidence: 99%

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Role for Ribosome-Associated Complex and Stress-Seventy subfamily B (RAC-Ssb) in integral membrane protein translation

Acosta-Sampson

Döring

Lin

et al. 2017

Preprint

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Targeting of most integral membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle (SRP), which recognizes a hydrophobic signal sequence near the protein N-terminus. Proper folding of these proteins is monitored by the unfolded protein response, and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transmembrane helix–the signal sequence recognized by SRP–is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these integral membrane proteins can divert the nascent protein chains to the ribosome-associated complex (RAC) and Stress-Seventy Subfamily B (Ssb) chaperones. We also show that quality control of integral membrane proteins by RAC-Ssb couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Role for Ribosome-Associated Complex and Stress-Seventy subfamily B (RAC-Ssb) in integral membrane protein translation

Acosta-Sampson

Döring

Lin

et al. 2017

Preprint

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“…These observations suggest that sr7575 does not cause the widespread protein unfolding that is typical of strong ER stress aggravators such as DTT and tunicamycin. Specific ER stress can be induced, for example, by expressing topologically abnormal ERAD-targeted integral membrane proteins without inducing the canonical UPR pathway in yeast (53).…”

Section: Discussionmentioning

confidence: 99%

The Toxicity of a Novel Antifungal Compound Is Modulated by Endoplasmic Reticulum-Associated Protein Degradation Components

Raj

Krishnan

Askew

et al. 2016

Antimicrob Agents Chemother

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iIn a search for new antifungal compounds, we screened a library of 4,454 chemicals for toxicity against the human fungal pathogen Aspergillus fumigatus. We identified sr7575, a molecule that inhibits growth of the evolutionary distant fungi A. fumigatus, Cryptococcus neoformans, Candida albicans, and Saccharomyces cerevisiae but lacks acute toxicity for mammalian cells. To gain insight into the mode of inhibition, sr7575 was screened against 4,885 S. cerevisiae mutants from the systematic collection of haploid deletion strains and 977 barcoded haploid DAmP (decreased abundance by mRNA perturbation) strains in which the function of essential genes was perturbed by the introduction of a drug resistance cassette downstream of the coding sequence region. Comparisons with previously published chemogenomic screens revealed that the set of mutants conferring sensitivity to sr7575 was strikingly narrow, affecting components of the endoplasmic reticulum-associated protein degradation (ERAD) stress response and the ER membrane protein complex (EMC). ERAD-deficient mutants were hypersensitive to sr7575 in both S. cerevisiae and A. fumigatus, indicating a conserved mechanism of growth inhibition between yeast and filamentous fungi. Although the unfolded protein response (UPR) is linked to ERAD regulation, sr7575 did not trigger the UPR in A. fumigatus and UPR mutants showed no enhanced sensitivity to the compound. The data from this chemogenomic analysis demonstrate that sr7575 exerts its antifungal activity by disrupting ER protein quality control in a manner that requires ERAD intervention but bypasses the need for the canonical UPR. ER protein quality control is thus a specific vulnerability of fungal organisms that might be exploited for antifungal drug development.

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“…2D). UPR activation has been observed during expression of membrane protein mutants that destabilize the native structure or during overproduction of specific membrane proteins for crystallographic purposes (43,44). Although Opt was not misfolded ( Fig.…”

Section: Codon-optimized Cdt-1 Activates the Unfolded Protein Responsementioning

confidence: 99%

“…Our results suggest that RAC-Ssb may be responsible for this role in the context of at least the MFS class of membrane proteins. When rapidly translated membrane proteins, such as Opt, are highly expressed, RAC-Ssb interactions with these nascent chains and the subsequent degradation of a subset of these polypeptides could overwhelm the basal level of protein quality control in cells, which would increase the proteasomal load and trigger cell stress and possibly the UPR (44,71). Notably, transporters were highly enriched in the subset of proteins identified to be toxic in a genome-wide analysis of protein overexpression in yeast (74), consistent with our model for RAC-Ssb function and the relationship between OPT expression and cell growth (supplemental Fig.…”

Section: Rac-ssb and Integral Membrane Protein Translationmentioning

confidence: 99%

Role for ribosome-associated complex and stress-seventy subfamily B (RAC-Ssb) in integral membrane protein translation

Acosta-Sampson

Döring

Lin

et al. 2017

Journal of Biological Chemistry

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Targeting of most integral membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle, which recognizes a hydrophobic signal sequence near the protein N terminus. Proper folding of these proteins is monitored by the unfolded protein response and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transmembrane helix, the signal sequence recognized by the signal recognition particle, is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these integral membrane proteins can divert the nascent protein chains to the ribosome-associated complex and stress-seventy subfamily B chaperones. We also show that quality control of integral membrane proteins by ribosome-associated complex-stress-seventy subfamily B couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology.

show abstract

Expression of three topologically distinct membrane proteins elicits unique stress response pathways in the yeastSaccharomyces cerevisiae

Abstract: Buck TM, Jordan R, Lyons-Weiler J, Adelman JL, Needham PG, Kleyman TR, Brodsky JL. Expression of three topologically distinct membrane proteins elicits unique stress response pathways in the yeast

Cited by 10 publications

References 171 publications

Role for Ribosome-Associated Complex and Stress-Seventy subfamily B (RAC-Ssb) in integral membrane protein translation

Role for Ribosome-Associated Complex and Stress-Seventy subfamily B (RAC-Ssb) in integral membrane protein translation

The Toxicity of a Novel Antifungal Compound Is Modulated by Endoplasmic Reticulum-Associated Protein Degradation Components

Role for ribosome-associated complex and stress-seventy subfamily B (RAC-Ssb) in integral membrane protein translation

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