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Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome

Lateral ribosomal stalk is responsible for binding and recruiting translation factors during protein synthesis. The eukaryotic stalk consists of one P0 protein with two copies of P1•P2 heterodimers to form a P0(P1•P2)(2) pentameric P-complex. Here, we have solved the structure of full-length P1•P2 b...

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Autores principales: Lee, Ka-Ming, Yusa, Kazuyuki, Chu, Lai-On, Yu, Conny Wing-Heng, Oono, Moe, Miyoshi, Tomohiro, Ito, Kosuke, Shaw, Pang-Chui, Wong, Kam-Bo, Uchiumi, Toshio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3794596/
https://www.ncbi.nlm.nih.gov/pubmed/23892290
http://dx.doi.org/10.1093/nar/gkt636
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author Lee, Ka-Ming
Yusa, Kazuyuki
Chu, Lai-On
Yu, Conny Wing-Heng
Oono, Moe
Miyoshi, Tomohiro
Ito, Kosuke
Shaw, Pang-Chui
Wong, Kam-Bo
Uchiumi, Toshio
author_facet Lee, Ka-Ming
Yusa, Kazuyuki
Chu, Lai-On
Yu, Conny Wing-Heng
Oono, Moe
Miyoshi, Tomohiro
Ito, Kosuke
Shaw, Pang-Chui
Wong, Kam-Bo
Uchiumi, Toshio
author_sort Lee, Ka-Ming
collection PubMed
description Lateral ribosomal stalk is responsible for binding and recruiting translation factors during protein synthesis. The eukaryotic stalk consists of one P0 protein with two copies of P1•P2 heterodimers to form a P0(P1•P2)(2) pentameric P-complex. Here, we have solved the structure of full-length P1•P2 by nuclear magnetic resonance spectroscopy. P1 and P2 dimerize via their helical N-terminal domains, whereas the C-terminal tails of P1•P2 are unstructured and can extend up to ∼125 Å away from the dimerization domains. (15)N relaxation study reveals that the C-terminal tails are flexible, having a much faster internal mobility than the N-terminal domains. Replacement of prokaryotic L10(L7/L12)(4)/L11 by eukaryotic P0(P1•P2)(2)/eL12 rendered Escherichia coli ribosome, which is insensitive to trichosanthin (TCS), susceptible to depurination by TCS and the C-terminal tail was found to be responsible for this depurination. Truncation and insertion studies showed that depurination of hybrid ribosome is dependent on the length of the proline-alanine rich hinge region within the C-terminal tail. All together, we propose a model that recruitment of TCS to the sarcin-ricin loop required the flexible C-terminal tail, and the proline-alanine rich hinge region lengthens this C-terminal tail, allowing the tail to sweep around the ribosome to recruit TCS.
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spelling pubmed-37945962013-10-21 Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome Lee, Ka-Ming Yusa, Kazuyuki Chu, Lai-On Yu, Conny Wing-Heng Oono, Moe Miyoshi, Tomohiro Ito, Kosuke Shaw, Pang-Chui Wong, Kam-Bo Uchiumi, Toshio Nucleic Acids Res Structural Biology Lateral ribosomal stalk is responsible for binding and recruiting translation factors during protein synthesis. The eukaryotic stalk consists of one P0 protein with two copies of P1•P2 heterodimers to form a P0(P1•P2)(2) pentameric P-complex. Here, we have solved the structure of full-length P1•P2 by nuclear magnetic resonance spectroscopy. P1 and P2 dimerize via their helical N-terminal domains, whereas the C-terminal tails of P1•P2 are unstructured and can extend up to ∼125 Å away from the dimerization domains. (15)N relaxation study reveals that the C-terminal tails are flexible, having a much faster internal mobility than the N-terminal domains. Replacement of prokaryotic L10(L7/L12)(4)/L11 by eukaryotic P0(P1•P2)(2)/eL12 rendered Escherichia coli ribosome, which is insensitive to trichosanthin (TCS), susceptible to depurination by TCS and the C-terminal tail was found to be responsible for this depurination. Truncation and insertion studies showed that depurination of hybrid ribosome is dependent on the length of the proline-alanine rich hinge region within the C-terminal tail. All together, we propose a model that recruitment of TCS to the sarcin-ricin loop required the flexible C-terminal tail, and the proline-alanine rich hinge region lengthens this C-terminal tail, allowing the tail to sweep around the ribosome to recruit TCS. Oxford University Press 2013-10 2013-07-26 /pmc/articles/PMC3794596/ /pubmed/23892290 http://dx.doi.org/10.1093/nar/gkt636 Text en © The Author(s) 2013. Published by Oxford University Press. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Structural Biology
Lee, Ka-Ming
Yusa, Kazuyuki
Chu, Lai-On
Yu, Conny Wing-Heng
Oono, Moe
Miyoshi, Tomohiro
Ito, Kosuke
Shaw, Pang-Chui
Wong, Kam-Bo
Uchiumi, Toshio
Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome
title Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome
title_full Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome
title_fullStr Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome
title_full_unstemmed Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome
title_short Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome
title_sort solution structure of human p1•p2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3794596/
https://www.ncbi.nlm.nih.gov/pubmed/23892290
http://dx.doi.org/10.1093/nar/gkt636
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