Auxin Response Factor 6 and 8 LC-MS

ABSTRACT: The phytohormone auxin plays a pivotal role in promoting fruit initiation and growth upon fertilization in flowering plants. Upregulation of auxin signaling by genetic mutations or exogenous auxin treatment can induce seedless fruit formation from unpollinated ovaries, termed parthenocarpy. Recent studies suggested that the class A AUXIN RESPONSE FACTOR6 (ARF6) and ARF8 play dual functions by first inhibiting fruit initiation when complexed with unidentified corepressor IAA protein(s) before pollination, and later promoting fruit growth after fertilization as ARF dimers. However, whether and how posttranslational modification(s) regulate ARF6- and ARF8-mediated fruit growth were unknown. In this study, we reveal that both ARF6 and ARF8 are O-fucosylated in their middle region (MR) by SPINDLY (SPY), a novel nucleocytoplasmic protein O-fucosyltransferase, which catalyzes the addition of a fucose moiety to specific Ser/Thr residues of target proteins. Epistasis, biochemical and transcriptome analyses indicated that ARF6 and ARF8 are downstream of SPY, but ARF8 plays a more predominant role in parthenocarpic fruit growth. Intriguingly, two ARF6/8 interacting proteins, the co-repressor IAA9 and MED8, a subunit of the coactivator Mediator complex, were also O-fucosylated by SPY. Biochemical assays demonstrated that SPY-mediated O-fucosylation of these proteins reduced ARF-MED8 interaction, which led to enhanced transcription repression activity of the ARF6/8-IAA9 complex but impaired transactivation activities of ARF6/8. Our study unveils the role of protein O-fucosylation by SPY in attenuating auxin-triggered fruit growth through modulation of activities of key transcription factors, co-repressor and the coactivator MED complex.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Nicotiana Benthamiana Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Seedling, Leaf

SUBMITTER: Seamus Kelley  

LAB HEAD: Dr Jeffrey Shabanowitz

PROVIDER: PXD051232 | Pride | 2025-03-27

REPOSITORIES: Pride