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Translation elongation factor 1A is essential for regulation of the actin cytoskeleton and cell morphology

Nature Structural & Molecular Biology volume 12, pages 772–778 (2005)Cite this article

Abstract

The binding of eukaryotic translation elongation factor 1A (eEF1A) to actin is a noncanonical function that may link two distinct cellular processes, cytoskeleton organization and gene expression. Using the yeast Saccharomyces cerevisiae, we have established an in vivo assay that directly identifies specific regions and residues of eEF1A responsible for actin interactions and bundling. Using a unique genetic screen, we isolated a series of eEF1A mutants with reduced actin bundling activity. These mutations alter actin cytoskeleton organization but not translation, indicating that these are separate functions of eEF1A. This demonstrates for the first time a direct consequence of eEF1A on cytoskeletal organization in vivo and the physiological significance of this interaction.

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Figure 1: Residues 414–427 of domain III of eEF1A are essential for growth inhibition, actin disorganization and cell cycle phenotypes conferred by eEF1A overexpression.
Figure 2: The F422A domain III mutant of eEF1A partially suppresses the growth inhibition and actin disorganization conferred by eEF1A overexpression.
Figure 3: N305S and N329S point mutants of eEF1A-Ura3p do not confer the overexpression phenotypes and cluster on one face of the eEF1A.
Figure 4: N305S and N329S point mutants of eEF1A-Ura3p show modest differences in growth and no change in translation rates.
Figure 5: N305S and N329S eEF1A-Ura3p mutants are deficient in bundling actin in vitro and show differences in cell morphology and actin cytoskeletal organization in vivo.
Figure 6: N305S and N329S eEF1A-Ura3p mutant strains show significantly increased levels of soluble actin.

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Acknowledgements

The authors acknowledge the assistance of the Cancer Institute of New Jersey/Environmental and Occupational Health Sciences Institute Analytical Cytometry Image Analysis, the Robert Wood Johnson Medical School (RWJMS) DNA Core Facility sequencing laboratory and the fluorescence microscopy laboratory in the RWJMS Department of Pharmacology. This research was supported by a grant from the US National Institutes of Health (GM62789 to T.G.K.).

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  1. Department of Molecular Genetics, Microbiology & Immunology, University of Medicine and Dentistry of New Jersey–Robert Wood Johnson Medical School, Piscataway, 08854, New Jersey, USA

    Stephane R Gross & Terri Goss Kinzy

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Correspondence to Terri Goss Kinzy.

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Supplementary information

Supplementary Fig. 1

N305S and N329S eEF1A/Ura3p mutant do not show polyribosomes accumulation. (PDF 1118 kb)

Supplementary Table 1

S. cerevisiae strains used in this study. (PDF 97 kb)

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Gross, S., Kinzy, T. Translation elongation factor 1A is essential for regulation of the actin cytoskeleton and cell morphology. Nat Struct Mol Biol 12, 772–778 (2005). https://doi.org/10.1038/nsmb979

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