Our data are first to indicate that the individual subunits can exist separately from the eEF1B complex in cancer tissues and that disintegration of eEF1B could be an important sign of cancer development. Nuclear localization of Bγ both in normal and in cancer tissues suggests its previously unknown nucleus-specific role in human cells.

BackgroundThe constituents of stable multiprotein complexes are known to dissociate from the complex to play independent regulatory roles. The components of translation elongation complex eEF1H (eEF1A, eEF1Bα, eEF1Bβ, eEF1Bγ) were found overexpressed in different cancers. To gain the knowledge about novel cancer-related translational mechanisms we intended to reveal whether eEF1H exists as a single unit or independent subunits in different human cancers.MethodsThe changes in the expression level of every subunit of eEF1H in the human non-small-cell lung cancer tissues were examined. The localization of eEF1H subunits was assessed by immunohistochemistry methods, subcellular fractionation and confocal microscopy. The possibility of the interaction between the subunits was estimated by co-immunoprecipitation.ResultsThe level of eEF1Bβ expression was increased more than two-fold in 36%, eEF1Bγ in 28%, eEF1A in 20% and eEF1Bα in 8% of tumor specimens. The cancer-induced alterations in the subunits level were found to be uncoordinated, therefore the increase in the level of at least one subunit of eEF1H was observed in 52% of samples. Nuclear localization of eEF1Bβ in the cancer rather than distal normal looking tissues was found. In cancer tissue, eEF1A and eEF1Bα were not found in nuclei while all four subunits of eEF1H demonstrated both cytoplasmic and nuclear appearance in the lung carcinoma cell line A549. Unexpectedly, in the A549 nuclear fraction eEF1A lost the ability to interact with the eEF1B complex.ConclusionsThe results suggest independent functioning of some fraction of the eEF1H subunits in human tumors. The absence of eEF1A and eEF1B interplay in nuclei of A549 cells is a first evidence for non-translational role of nuclear-localized subunits of eEF1B. We conclude the appearance of the individual eEF1B subunits in tumors is a more general phenomenon than appreciated before and thus is a novel signal of cancer-related changes in translation apparatus.

Eukaryotic elongation factor 1 (eEF1) mediates the binding of aminoacyl-tRNA to the ribosome in GTP-dependent manner. eEF1 consists of four subunits: eEF1A, eEF1Ba, eEF1Bb and eEF1Bg. eEF1A has two different isoforms: eEF1A1 is present throughout development and is ubiquitously expressed with the exception of adult muscle, while eEF1A2 is developmentally regulated and expressed only in muscle cells and neurons. Expression of eEF1A1, eEF1A2, eEF1Ba, eEF1Bb and eEF1Bg genes was analyzed by Northern blot hybridization of a panel of brain tumor and normal brain tissue RNAs. Totally 23 glioblastoma and 10 normal brain samples were investigated. In gliomas, no meaningful difference in the mRNA content for the eEF1A1, eEF1Ba and eEF1Bg subunits as compared to normal brain tissues was found. However, we have observed approximately 2-fold decrease in the eEF1Bb mRNA expression in human gliomas as compared to normal human brain by Northern blot analysis. Besides, we have shown reduced level of the eEF1A2 mRNA expression in glioblastoma as compared to normal human glia.

Aim. To investigate protein level of all subunits of the eukaryotic elongation translation factor eEF1H (eEF1A, eEF1Bα, eEF1Bβand eEF1Bγ) in glial tumors of human brain in comparison with normal brain. Methods. The eEF1H components content has been investigated in human glioblastoma clinical samples by Western blot analysis. Results. To determine the eEF1Bα, eEF1Bβ and eEF1Bγ content, the polyclonal antibodies against all eEF1H subunits were obtained. The tendency of the eEF1Bγ protein level to increase in glioblastomas was observed. There were no significant differences in the eEF1A, eEF1Bα and eEF1Bβ protein contents. Conclusions. In the previous report we analysed the expression of all eEF1H subunits in human glial brain tumor on the mRNA level. This study showed that eEF1Bγ was overexpressed while no significant changes in other eEF1H subunits were observed. It suggests a possible function of eEFBg which is cancer-related and is not connected with the functioning of eEF1H complex in translation