The ASCC2 CUE domain in the ALKBH3–ASCC DNA repair complex recognizes adjacent ubiquitins in K63-linked polyubiquitin

Lombardi, Patrick M.; Haile, Sara; Rusanov, Timur; Rodell, Rebecca; Anoh, Rita; Baer, Julia; Burke, Kate A.; Gray, Lauren N.; Hacker, Abigail R.; Kebreau, Kayla R.; Ngandu, Christine K.; Orland, Hannah A.; Osei-Asante, Emmanuella; Schmelyun, Dhane P.; Shorb, Devin E.; Syed, Shaheer H.; Veilleux, Julianna M.; Majumdar, Ananya; Mosammaparast, Nima; Wolberger, Cynthia

doi:10.1016/j.jbc.2021.101545

Cited by 7 publications

(6 citation statements)

References 28 publications

(119 reference statements)

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“…5). In the first step, the RQT is recruited to the ribosome queue after Hel2-dependent K63-linked uS10 polyubiquitination, likely via interaction with the CUE domain of the Cue3 subunit ( 7, 36, 37 ). After recruitment, RQT stably locks onto the 40S of the stalled/lead ribosome in the C1 conformation via the Slh1 NTC, placing the Slh1 CTC in a position primed to bind the 3’ region of the mRNA emerging from the ribosomal mRNA entry site.…”

Section: Resultsmentioning

confidence: 99%

Clearing of ribosome collisions by the ribosome quality control trigger complex RQT

Best¹,

Ikeuchi²,

Kater³

et al. 2022

Preprint

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After translational stalls, colliding eukaryotic ribosomes are cleared through dissociation into subunits by the ribosome quality control trigger complex, RQT, by an unknown mechanism. Here we show that RQT requires accessible mRNA and the presence of a neighboring ribosome. Cryo-EM of several RQT-ribosome complexes revealed the structural basis of splitting: RQT engages the 40S subunit of the lead ribosome and can switch between two conformations. We propose a mechanistic model in which the Slh1 helicase subunit of RQT applies a pulling force on the mRNA, causing destabilizing conformational changes of the 40S subunit. The collided ribosome functions as a ram or giant wedge, ultimately resulting in subunit dissociation. Our findings provide a first conceptual framework for a helicase driven ribosomal splitting mechanism.One-Sentence SummaryRQT clears collided ribosomes by pulling mRNA to trigger destabilizing conformational transitions for subunit dissociation.

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Section: Resultsmentioning

confidence: 99%

Clearing of ribosome collisions by the ribosome quality control trigger complex RQT

Best¹,

Ikeuchi²,

Kater³

et al. 2022

Preprint

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show abstract

“…In the first step, the RQT is recruited to the ribosome queue after Hel2-dependent K63-linked uS10 polyubiquitination via interaction with the CUE domain of the Cue3 subunit 12 , 22 , 46 , 47 . After recruitment, RQT stably locks onto the 40S of the stalled/lead ribosome in the C1 conformation via the Slh1 NTC, placing the Slh1 CTC in a position primed to bind the 3′ region of the mRNA emerging from the ribosomal mRNA entry site.…”

Section: Discussionmentioning

confidence: 99%

Structural basis for clearing of ribosome collisions by the RQT complex

Best¹,

Ikeuchi²,

Kater

et al. 2023

Nat Commun

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Translation of aberrant messenger RNAs can cause stalling of ribosomes resulting in ribosomal collisions. Collided ribosomes are specifically recognized to initiate stress responses and quality control pathways. Ribosome-associated quality control facilitates the degradation of incomplete translation products and requires dissociation of the stalled ribosomes. A central event is therefore the splitting of collided ribosomes by the ribosome quality control trigger complex, RQT, by an unknown mechanism. Here we show that RQT requires accessible mRNA and the presence of a neighboring ribosome. Cryogenic electron microscopy of RQT-ribosome complexes reveals that RQT engages the 40S subunit of the lead ribosome and can switch between two conformations. We propose that the Ski2-like helicase 1 (Slh1) subunit of RQT applies a pulling force on the mRNA, causing destabilizing conformational changes of the small ribosomal subunit, ultimately resulting in subunit dissociation. Our findings provide conceptual framework for a helicase-driven ribosomal splitting mechanism.

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“…It is also possible that the protein interactions of ASCC3 may be dynamically regulated by post-translational modifications or by the recruitment of subsets of factors to specific sub-cellular compartments. Both of the latter principles have been shown to play a role during ASCC3-related cellular processes 13 , 14 , 26 , 31 , 32 , 41 , 47 .…”

Section: Discussionmentioning

confidence: 99%

“…Originally, ASCC3 was discovered as a component of the human ASCC that additionally encompasses subunits ASCC1 (containing RNA-binding KH and RNA ligase-like domains 13 ), ASCC2 (containing a K63-linked ubiquitin chain-binding CUE domain 14 ) and activating signal co-integrator 1/thyroid receptor-interacting protein 4 (TRIP4) 15 . By associating with basal transcription factors 16 , nuclear receptors 16 , 17 and/or various co-activators 15 , 16 , 18 , 19 , ASCC is thought to establish distinct transcription co-activator complexes in response to different cellular conditions 16 , 18 .…”

Section: Introductionmentioning

confidence: 99%

Extended DNA threading through a dual-engine motor module of the activating signal co-integrator 1 complex

et al. 2023

Self Cite

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Activating signal co-integrator 1 complex (ASCC) subunit 3 (ASCC3) supports diverse genome maintenance and gene expression processes, and contains tandem Ski2-like NTPase/helicase cassettes crucial for these functions. Presently, the molecular mechanisms underlying ASCC3 helicase activity and regulation remain unresolved. We present cryogenic electron microscopy, DNA-protein cross-linking/mass spectrometry as well as in vitro and cellular functional analyses of the ASCC3-TRIP4 sub-module of ASCC. Unlike the related spliceosomal SNRNP200 RNA helicase, ASCC3 can thread substrates through both helicase cassettes. TRIP4 docks on ASCC3 via a zinc finger domain and stimulates the helicase by positioning an ASC-1 homology domain next to the C-terminal helicase cassette of ASCC3, likely supporting substrate engagement and assisting the DNA exit. TRIP4 binds ASCC3 mutually exclusively with the DNA/RNA dealkylase, ALKBH3, directing ASCC3 for specific processes. Our findings define ASCC3-TRIP4 as a tunable motor module of ASCC that encompasses two cooperating NTPase/helicase units functionally expanded by TRIP4.

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The ASCC2 CUE domain in the ALKBH3–ASCC DNA repair complex recognizes adjacent ubiquitins in K63-linked polyubiquitin

Cited by 7 publications

References 28 publications

Clearing of ribosome collisions by the ribosome quality control trigger complex RQT

Clearing of ribosome collisions by the ribosome quality control trigger complex RQT

Structural basis for clearing of ribosome collisions by the RQT complex

Extended DNA threading through a dual-engine motor module of the activating signal co-integrator 1 complex

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