Here, we show that the eukaryotic translation elongation factor 1 gamma (eEF1gamma) physically interacts with the RNA polymerase II (pol II) core subunit 3 (RPB3), both in isolation and in the context of the holo-enzyme. Importantly, eEF1gamma has been recently shown to bind Vimentin mRNA. By chromatin immunoprecipitation experiments, we demonstrate, for the first time, that eEF1³ is also physically present on the genomic locus corresponding to the promoter region of human Vimentin gene. The eEF1³ depletion causes the Vimentin protein to be incorrectly compartmentalised and to severely compromise cellular shape and mitochondria localisation. We demonstrate that eEF1³ partially colocalises with the mitochondrial marker Tom20 and that eEF1³ depletion increases mitochondrial superoxide generation as well as the total levels of carbonylated proteins. Finally, we hypothesise that eEF1gamma, in addition to its role in translation elongation complex, is involved in regulating Vimentin gene by contacting both pol II and the Vimentin promoter region and then shuttling/nursing the Vimentin mRNA from its gene locus to its appropriate cellular compartment for translation.
The eEF1gamma Subunit Contacts RNA Polymerase II and Binds Vimentin Promoter Region
Pisani C;Onori A;Di Certo MG;Strimpakos G;Mattei E;Passananti C
2010
Abstract
Here, we show that the eukaryotic translation elongation factor 1 gamma (eEF1gamma) physically interacts with the RNA polymerase II (pol II) core subunit 3 (RPB3), both in isolation and in the context of the holo-enzyme. Importantly, eEF1gamma has been recently shown to bind Vimentin mRNA. By chromatin immunoprecipitation experiments, we demonstrate, for the first time, that eEF1³ is also physically present on the genomic locus corresponding to the promoter region of human Vimentin gene. The eEF1³ depletion causes the Vimentin protein to be incorrectly compartmentalised and to severely compromise cellular shape and mitochondria localisation. We demonstrate that eEF1³ partially colocalises with the mitochondrial marker Tom20 and that eEF1³ depletion increases mitochondrial superoxide generation as well as the total levels of carbonylated proteins. Finally, we hypothesise that eEF1gamma, in addition to its role in translation elongation complex, is involved in regulating Vimentin gene by contacting both pol II and the Vimentin promoter region and then shuttling/nursing the Vimentin mRNA from its gene locus to its appropriate cellular compartment for translation.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
