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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...

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Detalles Bibliográficos
Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
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
Descripción
Sumario: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.