A TOR (target of rapamycin) and nutritional phosphoproteome of fission yeast reveals novel targets in networks conserved in humans
Lenka Halova,
David Cobley,
Mirita Franz-Wachtel,
Tingting Wang,
Kaitlin R. Morrison,
Karsten Krug,
Nicolas Nalpas,
Boris Maček,
Iain M. Hagan,
Sean J. Humphrey,
Janni Petersen
Affiliations
Lenka Halova
Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK
David Cobley
Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Mirita Franz-Wachtel
Proteome Center Tuebingen, University of Tuebingen, Auf der Morgenstelle 15, 72076 Tuebingen, Germany
Tingting Wang
Flinders Health and Medical Research Institute, Flinders Centre for Innovation in Cancer, Flinders University, Adelaide, South Australia 5042, Australia
Kaitlin R. Morrison
Flinders Health and Medical Research Institute, Flinders Centre for Innovation in Cancer, Flinders University, Adelaide, South Australia 5042, Australia
Karsten Krug
Proteome Center Tuebingen, University of Tuebingen, Auf der Morgenstelle 15, 72076 Tuebingen, Germany
Nicolas Nalpas
Proteome Center Tuebingen, University of Tuebingen, Auf der Morgenstelle 15, 72076 Tuebingen, Germany
Boris Maček
Proteome Center Tuebingen, University of Tuebingen, Auf der Morgenstelle 15, 72076 Tuebingen, Germany
Iain M. Hagan
Cancer Research UK Manchester Institute, Alderley Park, Macclesfield SK10 4TG, UK
Sean J. Humphrey
Charles Perkins Centre, School of Life and Environmental Sciences, The University of Sydney, Camperdown, New South Wales, Australia
Janni Petersen
Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Fluctuations in TOR, AMPK and MAP-kinase signalling maintain cellular homeostasis and coordinate growth and division with environmental context. We have applied quantitative, SILAC mass spectrometry to map TOR and nutrient-controlled signalling in the fission yeast Schizosaccharomyces pombe. Phosphorylation levels at more than 1000 sites were altered following nitrogen stress or Torin1 inhibition of the TORC1 and TORC2 networks that comprise TOR signalling. One hundred and thirty of these sites were regulated by both perturbations, and the majority of these (119) new targets have not previously been linked to either nutritional or TOR control in either yeasts or humans. Elimination of AMPK inhibition of TORC1, by removal of AMPKα (ssp2::ura4+), identified phosphosites where nitrogen stress-induced changes were independent of TOR control. Using a yeast strain with an ATP analogue-sensitized Cdc2 kinase, we excluded sites that were changed as an indirect consequence of mitotic control modulation by nitrogen stress or TOR signalling. Nutritional control of gene expression was reflected in multiple targets in RNA metabolism, while significant modulation of actin cytoskeletal components points to adaptations in morphogenesis and cell integrity networks. Reduced phosphorylation of the MAPKK Byr1, at a site whose human equivalent controls docking between MEK and ERK, prevented sexual differentiation when resources were sparse but not eliminated.
