The R2TP is a recently identified Hsp90 co-chaperone, composed of four proteins as follows: Pih1D1, RPAP3, and the AAA(+)-ATPases RUVBL1 and RUVBL2. In mammals, the R2TP is involved in the biogenesis of cellular machineries such as RNA polymerases, small nucleolar ribonucleoparticles and phosphatidylinositol 3-kinase-related kinases. Here, we characterize the spaghetti (spag) gene of Drosophila, the homolog of human RPAP3. This gene plays an essential function during Drosophila development. We show that Spag protein binds Drosophila orthologs of R2TP components and Hsp90, like its yeast counterpart. Unexpectedly, Spag also interacts and stimulates the chaperone activity of Hsp70. Using null mutants and flies with inducible RNAi, we show that spaghetti is necessary for the stabilization of snoRNP core proteins and target of rapamycin activity and likely the assembly of RNA polymerase II. This work highlights the strong conservation of both the HSP90/R2TP system and its clients and further shows that Spag, unlike Saccharomyces cerevisiae Tah1, performs essential functions in metazoans. Interaction of Spag with both Hsp70 and Hsp90 suggests a model whereby R2TP would accompany clients from Hsp70 to Hsp90 to facilitate their assembly into macromolecular complexes.
Drosophila Spag is the homolog of RNA polymerase II-associated protein 3 (RPAP3) and recruits the heat shock proteins 70 and 90 (Hsp70 and Hsp90) during the assembly of cellular machineries
Benbahouche Nel, H.; Iliopoulos, I.; Torok, I.; Marhold, J.; Henri, J.; Kajava, A. V.; Farkas, R.; Kempf, T.; Schnolzer, M.; Meyer, P.; Kiss, I.; Bertrand*, E.; Mechler*, B. M.; Pradet-Balade*, B.
J Biol Chem
2014-02-28 / vol 289 / pages 6236-47
1083-351X (Electronic) 0021-9258 (Linking)
Humans; Animals; *Models, Biological; Anti-Bacterial Agents/pharmacology; Carrier Proteins/genetics/metabolism; Drosophila melanogaster; Drosophila Proteins/genetics/*metabolism; Heat-Shock Proteins/genetics/*metabolism; Hsp70; HSP70 Heat-Shock Proteins/genetics/*metabolism; Hsp90; Molecular Chaperones/genetics/*metabolism; Protein Assembly; R2TP Complex; Ribonucleoproteins, Small Nucleolar/genetics/*metabolism; RNA Polymerase II; RNA Polymerase II/genetics/metabolism; Rpap3; Saccharomyces cerevisiae Proteins/genetics/metabolism; Saccharomyces cerevisiae/genetics/metabolism; Sirolimus/pharmacology; Small Nucleolar RNA (snoRNA); Spaghetti Gene; TOR Complex (TORC)