Functional Characterization of Target of Rapamycin (TOR) Signalling in Physcomitrella

ABSTRACT: Target of rapamycin (TOR) is a structurally and functionally conserved protein kinase and an important signalling hub in eukaryotes. The moss Physcomitrella (Physcomitrium patens) is a model organism for plant physiology, development, and evolution. However, little is known about TOR signalling in non-vascular plants, including Physcomitrella. Here, we report the effects of inhibiting TOR signalling in Physcomitrella. We identified and characterized Physcomitrella 12-kD FK506-binding protein (FKBP12), which binds TOR in the presence of rapamycin. Whereas the growth of wild-type protonema is unaffected by rapamycin treatment, overexpressing endogenous FKBP12 rendered the plant susceptible to the drug in a dose-dependent manner. Likewise, protonema growth was inhibited when the TOR-specific ATP-competitive inhibitor AZD8055 was present in the culture. We show that rapamycin and AZD8055 have pleiotropic effects, as they delay cell cycle progression and development, induce chlorosis, inhibit photosynthesis, and alter total protein content. Additionally, we identified and characterized PpTOR, PpLST8, and PpRAPTOR, key components of TOR complex 1 (TORC1), whereas RICTOR and mSIN1 of TORC2 are absent from the moss genome. We found that PpLst8 substitutes its homolog in yeast to allow cell growth. Physcomitrella mutants were generated with a conditional downregulation of PpTOR, PpLST8, and PpRAPTOR, respectively. These mutants were impaired in growth. Finally, we show TOR-dependent phosphorylation of a well-known TOR phospho-target, the ribosomal protein RPS6, via LC-MS/MS. Collectively, our results suggest that Physcomitrella growth and development is positively controlled by a conserved TOR kinase. We suggest to further dissect TOR signalling in Physcomitrella in order to elucidate signalling integration via TORC1 in plants.

INSTRUMENT(S):

ORGANISM(S): Physcomitrella Physcomitrella Patens Subsp. Patens (moss)

SUBMITTER: Sebastian Hoernstein  

LAB HEAD: Pitter Huesgen

PROVIDER: PXD069889 | Pride | 2026-02-09

REPOSITORIES: Pride