Widespread reorganization of metabolic enzymes into reversible assemblies upon nutrient starvation.
Proteins are likely to organize into complexes that assemble and disassemble depending on cellular needs. When approximately 800 yeast strains expressing GFP-tagged proteins were grown to stationary phase, a surprising number of proteins involved in intermediary metabolism and stress response were observed to form punctate cytoplasmic foci. The formation of these discrete physical structures was confirmed by immunofluorescence and mass spectrometry of untagged proteins. The purine biosynthetic enzyme Ade4-GFP formed foci in the absence of adenine, and cycling between punctate and diffuse phenotypes could be controlled by adenine subtraction and addition. Similarly, glutamine synthetase (Gln1-GFP) foci cycled reversibly in the absence and presence of glucose. The structures were neither targeted for vacuolar or autophagosome degradation nor colocalized with P bodies or major organelles. Thus, upon nutrient depletion we observe widespread protein assemblies displaying nutrient-specific formation and dissolution.
Narayanaswamy
Rammohan
R
Department of Chemistry and Biochemistry, University of Texas, Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Austin, TX 78712-1064, USA.
Levy
Matthew
M
Tsechansky
Mark
M
Stovall
Gwendolyn M
GM
O'Connell
Jeremy D
JD
Mirrielees
Jennifer
J
Ellington
Andrew D
AD
Marcotte
Edward M
EM
D003593:Cytoplasm / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000096:biosynthesis; D013058:Mass Spectrometry; D009097:Multienzyme Complexes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000254:growth & development / Q000648:ultrastructure; D014617:Vacuoles / Q000201:enzymology
D009097:Multienzyme Complexes; D049452:Green Fluorescent Proteins