PT  - JOURNAL ARTICLE
AU  - Fuller, Gregory G.
AU  - Han, Ting
AU  - Freeberg, Mallory A.
AU  - Moresco, James J.
AU  - Yates, John R
AU  - Kim, John K.
TI  - RNA promotes phase separation of glycolysis enzymes into yeast G bodies in hypoxia
AID  - 10.1101/638650
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 638650
4099  - http://biorxiv.org/content/early/2019/05/17/638650.short
4100  - http://biorxiv.org/content/early/2019/05/17/638650.full
AB  - In hypoxic stress conditions, glycolysis enzymes assemble into singular cytoplasmic granules called glycolytic (G) bodies. Formation of G bodies in yeast is correlated with increased glucose consumption and cell survival. However, the physical properties and organizing principles that define G body formation are unclear. We demonstrate that glycolysis enzymes are non-canonical RNA binding proteins, sharing many common mRNA substrates that are also integral constituents of G bodies. Tethering a G body component, the beta subunit of the yeast phosphofructokinase, Pfk2, to nonspecific endoribonucleases reveals that RNA nucleates G body formation and subsequent maintenance of G body structural integrity. Consistent with a phase separation mechanism of G body formation, recruitment of glycolysis enzymes to G bodies relies on multivalent homotypic and heterotypic interactions. Furthermore, G bodies can fuse in live cells and are largely insensitive to 1,6-hexanediol treatment, consistent with a hydrogel-like state in its composition. Taken together, our results elucidate the biophysical nature of G bodies and demonstrate that RNA nucleates phase separation of the glycolysis machinery in response to hypoxic stress.