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RqcH and RqcP catal...
RqcH and RqcP catalyze processive poly-alanine synthesis in a reconstituted ribosome-associated quality control system
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- Takada, Hiraku (author)
- Umeå University,Umeå universitet,Molekylär Infektionsmedicin, Sverige (MIMS),Institutionen för molekylärbiologi (Medicinska fakulteten),Faculty of Life Sciences, Kyoto Sangyo University, Kyoto, Japan
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- Crowe-Mcauliffe, Caillan (author)
- Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany
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- Polte, Christine (author)
- Konstantinov of Natl. Research Centre kurchatov Institute, Gatchina, Russian Federation; Russian Research Institute of Hematology and Transfusiology of FMBA, Saint Petersburg, Russian Federation,University of Hamburg
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- Sidorova, Zhanna Yu (author)
- Russian Research Center 'Kurchatov Institute',Russian Research Institute of Hematology and Transfusiology
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- Murina, Victoriia (author)
- Umeå University,Umeå universitet,Institutionen för molekylärbiologi (Medicinska fakulteten),Molekylär Infektionsmedicin, Sverige (MIMS)
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- Atkinson, Gemma C (author)
- Konstantinov of Natl. Research Centre kurchatov Institute, Gatchina, Russian Federation; National Research Centre "kurchatov Institute", Moscow, Russian Federation; Peter the Great St. Petersburg Polytechnic University, Saint Petersburg, Russian Federation
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- Konevega, Andrey L (author)
- Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany,Russian Research Center 'Kurchatov Institute',St. Petersburg State Polytechnical University
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- Ignatova, Zoya (author)
- Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany,Hamburg University of Technology
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- Wilson, Daniel N (author)
- Department of Experimental Medical Science, Lund University, Lund, Sweden; University of Tartu, Institute of Technology, Tartu, Estonia,University of Hamburg
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- Hauryliuk, Vasili, 1980- (author)
- Umeå University,Lund University,Lunds universitet,Umeå universitet,Institutionen för molekylärbiologi (Medicinska fakulteten),Molekylär Infektionsmedicin, Sverige (MIMS),Department of Experimental Medical Science, Lund University, 221 00 Lund, Sweden, University of Tartu, Institute of Technology, Tartu, Estonia,Molekylär enzymologi,Forskargrupper vid Lunds universitet,Molecular Enzymology,Lund University Research Groups
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(creator_code:org_t)
- 2021-07-13
- 2021
- English.
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In: Nucleic Acids Research. - : Oxford University Press. - 0305-1048 .- 1362-4962. ; 49:14, s. 8355-8369
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Abstract
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- In the cell, stalled ribosomes are rescued through ribosome-associated protein quality-control (RQC) pathways. After splitting of the stalled ribosome, a C-terminal polyalanine 'tail' is added to the unfinished polypeptide attached to the tRNA on the 50S ribosomal subunit. In Bacillus subtilis, polyalanine tailing is catalyzed by the NEMF family protein RqcH, in cooperation with RqcP. However, the mechanistic details of this process remain unclear. Here we demonstrate that RqcH is responsible for tRNAAla selection during RQC elongation, whereas RqcP lacks any tRNA specificity. The ribosomal protein uL11 is crucial for RqcH, but not RqcP, recruitment to the 50S subunit, and B. subtilis lacking uL11 are RQC-deficient. Through mutational mapping, we identify critical residues within RqcH and RqcP that are important for interaction with the P-site tRNA and/or the 50S subunit. Additionally, we have reconstituted polyalanine-tailing in vitro and can demonstrate that RqcH and RqcP are necessary and sufficient for processivity in a minimal system. Moreover, the in vitro reconstituted system recapitulates our in vivo findings by reproducing the importance of conserved residues of RqcH and RqcP for functionality. Collectively, our findings provide mechanistic insight into the role of RqcH and RqcP in the bacterial RQC pathway.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
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- ref (subject category)
- art (subject category)
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- By the author/editor
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Takada, Hiraku
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Crowe-Mcauliffe, ...
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Polte, Christine
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Sidorova, Zhanna ...
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Murina, Victorii ...
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Atkinson, Gemma ...
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show more...
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Konevega, Andrey ...
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Ignatova, Zoya
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Wilson, Daniel N
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Hauryliuk, Vasil ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
- Articles in the publication
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Nucleic Acids Re ...
- By the university
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Umeå University
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Lund University