Interaction of Rho1p target Bni1p with F-actin-binding elongation factor 1α: implication in Rho1p-regulated reorganization of the actin cytoskeleton in Saccharomyces cerevisiae

Abstract

The RHO1 gene encodes a homolog of mammalian RhoA small G protein in the yeast Saccharomyces cerevisiae. We have shown that Bni1p is one of the downstream targets of Rho1p and regulates reorganization of the actin cytoskeleton through the interaction with profilin, an actin monomer-binding protein. A Bni1p-binding protein was affinity purified from the yeast cytosol fraction and was identified to be Tef1p/Tef2p, translation elongation factor 1α (EF1α). EF1α is an essential component of the protein synthetic machinery and also possesses the actin filament (F-actin)-binding and -bundling activities. EF1α bound to the 186 amino acids region of Bni1p, located between the FH1 domain, the proline-rich profilin-binding domain, and the FH2 domain, of which function is not known. The binding of Bni1p to EF1α inhibited its F-actin-binding and -bundling activities. The BNI1 gene deleted in the EF1α-binding region did not suppress the bni1 bnr1 mutation in which the actin organization was impaired. These results suggest that the Rho1p–Bni1p system regulates reorganization of the actin cytoskeleton through the interaction with both EF1α and profilin.