Tetrahymena eukaryotic translation elongation factor 1A (eEF1A) bundles filamentous actin through dimer formation

J Biochem. 2006 Sep;140(3):393-9. doi: 10.1093/jb/mvj169. Epub 2006 Jul 27.

Abstract

Eukaryotic translation elongation factor 1A (eEF1A) is known to be a multifunctional protein. In Tetrahymena, eEF1A is localized to the division furrow and has the character to bundle filamentous actin (F-actin). eEF1A binds F-actin and the ratio of eEF1A and actin is approximately 1:1 (Kurasawa et al., 1996). In this study, we revealed that eEF1A itself exists as monomer and dimer, using gel filtration column chromatography. Next, eEF1A monomer and eEF1A dimer were separated using gel filtration column, and their interaction with F-actin was examined with cosedimentation assay and electron microscopy. In the absence of Ca2+/calmodulin (CaM), eEF1A dimer bundled F-actin and coprecipitated with F-actin at low-speed centrifugation, but eEF1A monomer did not. In the presence of Ca2+/CaM, eEF1A monomer increased, while dimer decreased. To examine that Ca2+/CaM alters eEF1A dimer into monomer and inhibits bundle formation of F-actin, Ca2+/CaM was added to F-actin bundles formed by eEF1A dimer. Ca2+/CaM separated eEF1A dimer to monomer, loosened F-actin bundles and then dispersed actin filaments. Simultaneously, Ca2+/CaM/ eEF1A monomer complexes were dissociated from actin filaments. Therefore, Ca2+/CaM reversibly regulates the F-actin bundling activity of eEF1A.

Publication types

  • Comparative Study

MeSH terms

  • Actins / metabolism*
  • Animals
  • Calmodulin / metabolism
  • Chromatography, Gel
  • Dimerization
  • Electrophoresis, Polyacrylamide Gel
  • Immunoblotting
  • Microscopy, Electron
  • Peptide Elongation Factor 1 / metabolism*
  • Tetrahymena / metabolism*

Substances

  • Actins
  • Calmodulin
  • Peptide Elongation Factor 1