Cryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG8

The PI3K-related kinase (PIKK) SMG1 monitors the progression of metazoan nonsense-mediated mRNA decay (NMD) by phosphorylating the RNA helicase UPF1. Previous work has shown that the activity of SMG1 is impaired by small molecule inhibitors, is reduced by the SMG1 interactors SMG8 and SMG9, and is d...

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Autores principales:	Lukas M Langer, Fabien Bonneau, Yair Gat, Elena Conti
Formato:	article
Lenguaje:	EN
Publicado:	eLife Sciences Publications Ltd 2021
Materias:	nonsense-mediated mRNA decay PIKK mRNA surveillance Medicine R Science Q Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/7f597ec2a3aa4b7b9ddb0d4dcef86fd1
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spelling	oai:doaj.org-article:7f597ec2a3aa4b7b9ddb0d4dcef86fd12021-11-15T09:43:20ZCryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG810.7554/eLife.723532050-084Xe72353https://doaj.org/article/7f597ec2a3aa4b7b9ddb0d4dcef86fd12021-10-01T00:00:00Zhttps://elifesciences.org/articles/72353https://doaj.org/toc/2050-084XThe PI3K-related kinase (PIKK) SMG1 monitors the progression of metazoan nonsense-mediated mRNA decay (NMD) by phosphorylating the RNA helicase UPF1. Previous work has shown that the activity of SMG1 is impaired by small molecule inhibitors, is reduced by the SMG1 interactors SMG8 and SMG9, and is downregulated by the so-called SMG1 insertion domain. However, the molecular basis for this complex regulatory network has remained elusive. Here, we present cryo-electron microscopy reconstructions of human SMG1-9 and SMG1-8-9 complexes bound to either a SMG1 inhibitor or a non-hydrolyzable ATP analog at overall resolutions ranging from 2.8 to 3.6 Å. These structures reveal the basis with which a small molecule inhibitor preferentially targets SMG1 over other PIKKs. By comparison with our previously reported substrate-bound structure (Langer et al.,2020), we show that the SMG1 insertion domain can exert an autoinhibitory function by directly blocking the substrate-binding path as well as overall access to the SMG1 kinase active site. Together with biochemical analysis, our data indicate that SMG1 autoinhibition is stabilized by the presence of SMG8. Our results explain the specific inhibition of SMG1 by an ATP-competitive small molecule, provide insights into regulation of its kinase activity within the NMD pathway, and expand the understanding of PIKK regulatory mechanisms in general.Lukas M LangerFabien BonneauYair GatElena ContieLife Sciences Publications Ltdarticlenonsense-mediated mRNA decayPIKKmRNA surveillanceMedicineRScienceQBiology (General)QH301-705.5ENeLife, Vol 10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	nonsense-mediated mRNA decay PIKK mRNA surveillance Medicine R Science Q Biology (General) QH301-705.5
spellingShingle	nonsense-mediated mRNA decay PIKK mRNA surveillance Medicine R Science Q Biology (General) QH301-705.5 Lukas M Langer Fabien Bonneau Yair Gat Elena Conti Cryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG8
description	The PI3K-related kinase (PIKK) SMG1 monitors the progression of metazoan nonsense-mediated mRNA decay (NMD) by phosphorylating the RNA helicase UPF1. Previous work has shown that the activity of SMG1 is impaired by small molecule inhibitors, is reduced by the SMG1 interactors SMG8 and SMG9, and is downregulated by the so-called SMG1 insertion domain. However, the molecular basis for this complex regulatory network has remained elusive. Here, we present cryo-electron microscopy reconstructions of human SMG1-9 and SMG1-8-9 complexes bound to either a SMG1 inhibitor or a non-hydrolyzable ATP analog at overall resolutions ranging from 2.8 to 3.6 Å. These structures reveal the basis with which a small molecule inhibitor preferentially targets SMG1 over other PIKKs. By comparison with our previously reported substrate-bound structure (Langer et al.,2020), we show that the SMG1 insertion domain can exert an autoinhibitory function by directly blocking the substrate-binding path as well as overall access to the SMG1 kinase active site. Together with biochemical analysis, our data indicate that SMG1 autoinhibition is stabilized by the presence of SMG8. Our results explain the specific inhibition of SMG1 by an ATP-competitive small molecule, provide insights into regulation of its kinase activity within the NMD pathway, and expand the understanding of PIKK regulatory mechanisms in general.
format	article
author	Lukas M Langer Fabien Bonneau Yair Gat Elena Conti
author_facet	Lukas M Langer Fabien Bonneau Yair Gat Elena Conti
author_sort	Lukas M Langer
title	Cryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG8
title_short	Cryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG8
title_full	Cryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG8
title_fullStr	Cryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG8
title_full_unstemmed	Cryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG8
title_sort	cryo-em reconstructions of inhibitor-bound smg1 kinase reveal an autoinhibitory state dependent on smg8
publisher	eLife Sciences Publications Ltd
publishDate	2021
url	https://doaj.org/article/7f597ec2a3aa4b7b9ddb0d4dcef86fd1
work_keys_str_mv	AT lukasmlanger cryoemreconstructionsofinhibitorboundsmg1kinaserevealanautoinhibitorystatedependentonsmg8 AT fabienbonneau cryoemreconstructionsofinhibitorboundsmg1kinaserevealanautoinhibitorystatedependentonsmg8 AT yairgat cryoemreconstructionsofinhibitorboundsmg1kinaserevealanautoinhibitorystatedependentonsmg8 AT elenaconti cryoemreconstructionsofinhibitorboundsmg1kinaserevealanautoinhibitorystatedependentonsmg8
_version_	1718428505104973824

Cryo-EM reconstructions of inhibitor-bound SMG1 kinase reveal an autoinhibitory state dependent on SMG8

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