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Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY
SPINDLY (SPY) in Arabidopsis thaliana is a novel nucleocytoplasmic protein O-fucosyltransferase (POFUT), which regulates diverse developmental processes. Sequence analysis indicates that SPY is distinct from ER-localized POFUTs and contains N-terminal tetratricopeptide repeats (TPRs) and a C-termina...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10027727/ https://www.ncbi.nlm.nih.gov/pubmed/36941311 http://dx.doi.org/10.1038/s41467-023-37279-1 |
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author | Kumar, Shivesh Wang, Yan Zhou, Ye Dillard, Lucas Li, Fay-Wei Sciandra, Carly A. Sui, Ning Zentella, Rodolfo Zahn, Emily Shabanowitz, Jeffrey Hunt, Donald F. Borgnia, Mario J. Bartesaghi, Alberto Sun, Tai-ping Zhou, Pei |
author_facet | Kumar, Shivesh Wang, Yan Zhou, Ye Dillard, Lucas Li, Fay-Wei Sciandra, Carly A. Sui, Ning Zentella, Rodolfo Zahn, Emily Shabanowitz, Jeffrey Hunt, Donald F. Borgnia, Mario J. Bartesaghi, Alberto Sun, Tai-ping Zhou, Pei |
author_sort | Kumar, Shivesh |
collection | PubMed |
description | SPINDLY (SPY) in Arabidopsis thaliana is a novel nucleocytoplasmic protein O-fucosyltransferase (POFUT), which regulates diverse developmental processes. Sequence analysis indicates that SPY is distinct from ER-localized POFUTs and contains N-terminal tetratricopeptide repeats (TPRs) and a C-terminal catalytic domain resembling the O-linked-N-acetylglucosamine (GlcNAc) transferases (OGTs). However, the structural feature that determines the distinct enzymatic selectivity of SPY remains unknown. Here we report the cryo-electron microscopy (cryo-EM) structure of SPY and its complex with GDP-fucose, revealing distinct active-site features enabling GDP-fucose instead of UDP-GlcNAc binding. SPY forms an antiparallel dimer instead of the X-shaped dimer in human OGT, and its catalytic domain interconverts among multiple conformations. Analysis of mass spectrometry, co-IP, fucosylation activity, and cryo-EM data further demonstrates that the N-terminal disordered peptide in SPY contains trans auto-fucosylation sites and inhibits the POFUT activity, whereas TPRs 1–5 dynamically regulate SPY activity by interfering with protein substrate binding. |
format | Online Article Text |
id | pubmed-10027727 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-100277272023-03-22 Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY Kumar, Shivesh Wang, Yan Zhou, Ye Dillard, Lucas Li, Fay-Wei Sciandra, Carly A. Sui, Ning Zentella, Rodolfo Zahn, Emily Shabanowitz, Jeffrey Hunt, Donald F. Borgnia, Mario J. Bartesaghi, Alberto Sun, Tai-ping Zhou, Pei Nat Commun Article SPINDLY (SPY) in Arabidopsis thaliana is a novel nucleocytoplasmic protein O-fucosyltransferase (POFUT), which regulates diverse developmental processes. Sequence analysis indicates that SPY is distinct from ER-localized POFUTs and contains N-terminal tetratricopeptide repeats (TPRs) and a C-terminal catalytic domain resembling the O-linked-N-acetylglucosamine (GlcNAc) transferases (OGTs). However, the structural feature that determines the distinct enzymatic selectivity of SPY remains unknown. Here we report the cryo-electron microscopy (cryo-EM) structure of SPY and its complex with GDP-fucose, revealing distinct active-site features enabling GDP-fucose instead of UDP-GlcNAc binding. SPY forms an antiparallel dimer instead of the X-shaped dimer in human OGT, and its catalytic domain interconverts among multiple conformations. Analysis of mass spectrometry, co-IP, fucosylation activity, and cryo-EM data further demonstrates that the N-terminal disordered peptide in SPY contains trans auto-fucosylation sites and inhibits the POFUT activity, whereas TPRs 1–5 dynamically regulate SPY activity by interfering with protein substrate binding. Nature Publishing Group UK 2023-03-20 /pmc/articles/PMC10027727/ /pubmed/36941311 http://dx.doi.org/10.1038/s41467-023-37279-1 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Kumar, Shivesh Wang, Yan Zhou, Ye Dillard, Lucas Li, Fay-Wei Sciandra, Carly A. Sui, Ning Zentella, Rodolfo Zahn, Emily Shabanowitz, Jeffrey Hunt, Donald F. Borgnia, Mario J. Bartesaghi, Alberto Sun, Tai-ping Zhou, Pei Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY |
title | Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY |
title_full | Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY |
title_fullStr | Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY |
title_full_unstemmed | Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY |
title_short | Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY |
title_sort | structure and dynamics of the arabidopsis o-fucosyltransferase spindly |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10027727/ https://www.ncbi.nlm.nih.gov/pubmed/36941311 http://dx.doi.org/10.1038/s41467-023-37279-1 |
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