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Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY.

Publication ,  Journal Article
Kumar, S; Wang, Y; Zhou, Y; Dillard, L; Li, F-W; Sciandra, CA; Sui, N; Zentella, R; Zahn, E; Shabanowitz, J; Hunt, DF; Borgnia, MJ; Sun, T-P ...
Published in: Nat Commun
March 20, 2023

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.

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Published In

Nat Commun

DOI

EISSN

2041-1723

Publication Date

March 20, 2023

Volume

14

Issue

1

Start / End Page

1538

Location

England

Related Subject Headings

  • Repressor Proteins
  • Fucosyltransferases
  • Fucose
  • Cryoelectron Microscopy
  • Arabidopsis Proteins
  • Arabidopsis
 

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Kumar, S., Wang, Y., Zhou, Y., Dillard, L., Li, F.-W., Sciandra, C. A., … Zhou, P. (2023). Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY. Nat Commun, 14(1), 1538. https://doi.org/10.1038/s41467-023-37279-1
Kumar, Shivesh, Yan Wang, Ye Zhou, Lucas Dillard, Fay-Wei Li, Carly A. Sciandra, Ning Sui, et al. “Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY.Nat Commun 14, no. 1 (March 20, 2023): 1538. https://doi.org/10.1038/s41467-023-37279-1.
Kumar S, Wang Y, Zhou Y, Dillard L, Li F-W, Sciandra CA, et al. Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY. Nat Commun. 2023 Mar 20;14(1):1538.
Kumar, Shivesh, et al. “Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY.Nat Commun, vol. 14, no. 1, Mar. 2023, p. 1538. Pubmed, doi:10.1038/s41467-023-37279-1.
Kumar S, Wang Y, Zhou Y, Dillard L, Li F-W, Sciandra CA, Sui N, Zentella R, Zahn E, Shabanowitz J, Hunt DF, Borgnia MJ, Bartesaghi A, Sun T-P, Zhou P. Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY. Nat Commun. 2023 Mar 20;14(1):1538.

Published In

Nat Commun

DOI

EISSN

2041-1723

Publication Date

March 20, 2023

Volume

14

Issue

1

Start / End Page

1538

Location

England

Related Subject Headings

  • Repressor Proteins
  • Fucosyltransferases
  • Fucose
  • Cryoelectron Microscopy
  • Arabidopsis Proteins
  • Arabidopsis