Cellular response to small molecules that selectively stall protein synthesis by the ribosome

Liaud, Nadège; Horlbeck, Max A.; Gilbert, Luke A.; Gjoni, Ketrin; Weissman, Jonathan S.; Cate, Jamie H. D.

doi:10.1371/journal.pgen.1008057

Cited by 33 publications

(33 citation statements)

References 50 publications

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“…We previously reported that an ortholog of yeast Slh1, ASCC3, is required for RQC 8 . In addition, a recent study suggested the involvement of ASCC3 and ASCC2 in co-translational quality control 20 . ASCC3 was originally identified as a component of the activating signal cointegrator 1 (ASC-1) complex, which consists of ASCC3, ASCC2, ASCC1, and TRIP4/ASC-1 (Fig.…”

Section: Analysis Of the Domains Of Znf598 Required For Rqc To Determentioning

confidence: 99%

“…We previously reported that ASCC3 is required for RQC induction 8 . A recent study showed that ASCC2 and ASCC3 bind to the ribosome and mitigate the toxic effects of stalling induced by PF8503 20 . The ATP-dependent helicase ASCC3 is an ortholog of Slh1(Rqt2) 8 , and the ubiquitin-binding protein ASCC2 has significant sequence homology to yeast Cue3(Rqt3), implying the existence of a human RQT (hRQT) complex.…”

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confidence: 99%

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Identification of a novel trigger complex that facilitates ribosome-associated quality control in mammalian cells

Hashimoto

Sugiyama

Yamazaki

et al. 2020

Sci Rep

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Ribosome stalling triggers the ribosome-associated quality control (RQC) pathway, which targets collided ribosomes and leads to subunit dissociation, followed by proteasomal degradation of the nascent peptide. In yeast, RQC is triggered by Hel2-dependent ubiquitination of uS10, followed by subunit dissociation mediated by the RQC-trigger (RQT) complex. In mammals, ZNF598-dependent ubiquitination of collided ribosomes is required for RQC, and activating signal cointegrator 3 (ASCC3), a component of the ASCC complex, facilitates RQC. However, the roles of other components and associated factors of the ASCC complex remain unknown. Here, we demonstrate that the human RQCtrigger (hRQT) complex, an ortholog of the yeast RQT complex, plays crucial roles in RQC. The hRQT complex is composed of ASCC3, ASCC2, and TRIP4, which are orthologs of the RNA helicase Slh1(Rqt2), ubiquitin-binding protein Cue3(Rqt3), and zinc-finger type protein yKR023W(Rqt4), respectively. The ATPase activity of ASCC3 and the ubiquitin-binding activity of ASCC2 are crucial for triggering RQC. Given the proposed function of the RQT complex in yeast, we propose that the hRQT complex recognizes the ubiquitinated stalled ribosome and induces subunit dissociation to facilitate RQC. Cells have evolved various quality control mechanisms to guarantee accurate gene expression 1-4. Ribosome stalling induces quality control mechanisms for mRNA, referred to as No-go decay (NGD) 5-7 , as well as for protein, referred to as ribosome-associated quality control (RQC) 8-11. RQC is conserved throughout species and consists primarily of four steps: (i) recognition of abnormal ribosome stalling; (ii) ubiquitination of specific residue(s) on the stalled ribosome; (iii) dissociation of ribosome into 40S and 60S subunits; and (iv) degradation of the nascent polypeptide on the 60S subunit 2,4. In the first step of RQC, the stalling of a ribosome at a specific sequence results in the formation of a di-ribosome (disome), which consists of the leading stalled ribosome and the following collided ribosome 5,6,10. Cryo-EM structural analysis has shown that the leading stalled ribosome is in the POST-state, with an empty A-site, whereas the colliding ribosome is in a rotated state with hybrid tRNAs 6,8,10. In the second step, the RING-type E3 ubiquitin ligase Hel2 recognizes the ribosome collision and ubiquitinates ribosomal protein uS10 (in yeast, at residue(s) K6/8) 8. In the third step, ubiquitinated ribosomes are dissociated into 40S subunits and 60S ribosome-nascent chain complexes (60S-RNCs), leading to subsequent RQC reactions. We recently proposed a model in which these ubiquitinated ribosomes are targeted by the RQT complex 8. The RQT complex is composed of three proteins: RNA helicase Slh1(Rqt2), ubiquitin-binding protein Cue3(Rqt3), and zinc-finger domain-containing protein yKR023W(Rqt4). The ubiquitin-binding activity of Cue3 and the ATPase activity of Slh1 are crucial for triggering RQC 8. After subunit dissociation, Rqc2 binds to tRNA at the subunit interface of 6...

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Section: Analysis Of the Domains Of Znf598 Required For Rqc To Determentioning

confidence: 99%

mentioning

confidence: 99%

Identification of a novel trigger complex that facilitates ribosome-associated quality control in mammalian cells

Hashimoto

Sugiyama

Yamazaki

et al. 2020

Sci Rep

View full text Add to dashboard Cite

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“…In addition, ASCC3 has been implicated in viral defense mechanisms 18 – 21 . Finally, a genome-wide CRISPRi screen showed that ASCC2 and ASCC3 are the two most potent modifiers of cell fitness in the presence of a translation inhibitor, suggesting that the proteins affect stalled ribosomes 22 . Indeed, ASCC3 has been suggested to resolve stalled ribosomes on poly-A sequences in a ribosome quality control pathway 23 – 25 .…”

Section: Introductionmentioning

confidence: 99%

The interaction of DNA repair factors ASCC2 and ASCC3 is affected by somatic cancer mutations

et al. 2020

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The ASCC3 subunit of the activating signal co-integrator complex is a dual-cassette Ski2-like nucleic acid helicase that provides single-stranded DNA for alkylation damage repair by the α-ketoglutarate-dependent dioxygenase AlkBH3. Other ASCC components integrate ASCC3/AlkBH3 into a complex DNA repair pathway. We mapped and structurally analyzed interacting ASCC2 and ASCC3 regions. The ASCC3 fragment comprises a central helical domain and terminal, extended arms that clasp the compact ASCC2 unit. ASCC2–ASCC3 interfaces are evolutionarily highly conserved and comprise a large number of residues affected by somatic cancer mutations. We quantified contributions of protein regions to the ASCC2–ASCC3 interaction, observing that changes found in cancers lead to reduced ASCC2–ASCC3 affinity. Functional dissection of ASCC3 revealed similar organization and regulation as in the spliceosomal RNA helicase Brr2. Our results delineate functional regions in an important DNA repair complex and suggest possible molecular disease principles.

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“…A comparison of the six CADs we screened vs. genome-scale screens (as opposed to targeted screens, e.g., involving the kinome) we and others performed with non-CADs shows the specificity of the CADs signature ( Figure 1C) (1)(2)(3)(4)(5). There were nineteen genes that scored on average an absolute value of at least 2.5 standard deviations away from the mean for the six CADs screens (hereafter referred to as pooled variance, PV) ( Figure 1C, Table S1).…”

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Sphingolipid Biosynthesis Inhibition As A Host Strategy Against Diverse Pathogens

Kumar

Park

Hart

et al. 2020

Preprint

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Chloroquine is an anti-malarial and immunosuppressant drug that has cationic amphipathic chemical properties. We performed genome-wide screens in human cells with chloroquine and several other widely used cationic amphipathic drugs (CADs) including the anti-depressants, sertraline (Zoloft) and fluoxetine (Prozac), the analgesic nortriptyline (Pamelor), the anti-arrhythmic amiodarone (Cordarone), and the anti-hypertensive verapamil (Calan) to characterize their molecular similarities and differences. Despite CADs having different disease indications but consistent with them sharing key chemical properties, we found CADs to have remarkably similar phenotypic profiles compared with non-CADs we and others have previously screened (1–5). The most significant genetic interaction for all CADs was the initiating step in sphingolipid biosynthesis catalyzed by serine palmitoyltransferase (SPT). A comparison of genome-wide screens performed with diverse pathogens from viruses, bacteria, plants, and parasites including Ebola (6), adeno-associated virus AAV2 (7), HIV (8), Rotavirus (9), Influenza A (10), Zika virus (11), Picornavirus (12), Exotoxin A (13), Cholera toxin (14), Type III secretion system and Shiga toxin (15, 16), Ricin toxin (17), and Toxoplasma gondii (18) showed SPT as a top common host factor and 80% overlap overall in top hits specifically with CADs. Potential sphingolipid-mediated mechanisms for the host response- and virulence-modulating effects of CADs involve autophagy and SERPINE1/PAI-1 (plasminogen activator inhibitor-1). Chloroquine has recently shown potential as an anti-viral agent for the novel coronavirus SARS-CoV-2, the causative agent of COVID-19 respiratory disease (19, 20). Our study demonstrates that numerous readily available drugs molecularly function highly similar to chloroquine, which suggests they might be considered for further pre-clinical investigation in the context of SARS-CoV-2. More generally, our work suggests the diverse pathogen mitigating potential of drugs that inhibit host sphingolipid biosynthesis such as CADs.Brief SummaryOur study demonstrates that numerous readily available drugs molecularly function highly similar to chloroquine, which suggests they might be considered for further pre-clinical investigation in the context of SARS-CoV-2.

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Cellular response to small molecules that selectively stall protein synthesis by the ribosome

Cited by 33 publications

References 50 publications

Identification of a novel trigger complex that facilitates ribosome-associated quality control in mammalian cells

Identification of a novel trigger complex that facilitates ribosome-associated quality control in mammalian cells

The interaction of DNA repair factors ASCC2 and ASCC3 is affected by somatic cancer mutations

Sphingolipid Biosynthesis Inhibition As A Host Strategy Against Diverse Pathogens

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