AKT/protein kinase B associates with beta-actin in the nucleus of melanoma cells

Author(s)

Coa, Larissa Leggieri ; Abreu, Thiago Ferreira ; Tashima, Alexandre Keiji ; Green, Janaina ; Pascon, Renata Castiglioni ; Vallim, Marcelo Afonso ; Machado-, Joel, Jr.

• Document type: Journal article
• Source: BIOSCIENCE REPORTS; v. 39, p. 15-pg., 2019-01-31.

Abstract

The serine-threonine kinase AKT/PKB is a critical regulator of various essential cellular processes, and dysregulation of AKT has been implicated in many diseases, including cancer. Despite AKT action is known to function mainly in the cytoplasm, AKT has been reported to translocate to the nucleus. However, very little is known about the mechanism required for the nuclear import of AKT as well as its function in this cellular compartment. In the present study, we characterized the presence of endogenous nuclear AKT in human melanoma cells and addressed the possible role of AKT by exploring its potential association with key interaction nuclear partners. Confocal and Western blot analyses showed that both phosphorylated and non-phosphorylated forms of AKT are present in melanoma cells nuclei. Using mass spectrometry in combination with protein-crosslinking and co-immunoprecipitation, we identified a series of putative protein partners of nuclear AKT, including heterogeneous nuclear ribonucleoprotein (hnRNP), cytoskeleton proteins beta-actin, gamma-actin, beta-actin-like 2 and vimentin. Confocal microscopy and biochemical analyses validated beta-actin as a new nuclear AKT-interacting partner. Cofilin and active RNA Polymerase II, two proteins that have been described to interact and work in concert with nuclear actin in transcription regulation, were also found associated with nuclear AKT. Overall, the present study uncovered a yet unrecognized nuclear coupling of AKT and provides insights into the involvement of AKT in the interaction network of nuclear actin. (AU)

• FAPESP's process: 12/19321-9 - Proteomic and peptidomic characterization of Brazilian spider venoms by mass spectrometry
• Grantee: Alexandre Keiji Tashima
• Support Opportunities: Regular Research Grants
• FAPESP's process: 11/50953-9 - Relevância funcional da quinase Akt/PKB nuclear na regulação da resistência ao anoikis em linhagens celulares de melanoma
• Grantee: Joel Machado Junior
• Support Opportunities: Regular Research Grants