Premium
Investigating the Control of the Actin Cytoskeleton by EF1α
Author(s) -
Colmer Sarah,
Karas Christina,
Scerbo Diego,
Gomez Maria,
Dunaway Stephen
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.550.5
Subject(s) - biology , schizosaccharomyces pombe , gene , actin , mutant , saccharomyces cerevisiae , eukaryotic translation elongation factor 1 alpha 1 , translation (biology) , microbiology and biotechnology , actin cytoskeleton , phalloidin , cytoskeleton , genetics , cell , ribosome , messenger rna , rna
Translation factor EF1α plays a critical role in protein synthesis, delivering aminoacyl tRNAs to the ribosome 1 . However, it has another less characterized function in regulating the actin cytoskeleton. EF1α binds actin directly or indirectly, as determined by co‐immunoprecipitation experiments 2 . Overexpression of EF1α in Schizosaccharomyces pombe results in aberrant cellular morphology 2 . EF1α homologs are potential cancer drug targets; the compound narciclasine, isolated from Amaryllidaceae , was shown to penetrate human melanoma cells, bind to eEF1A, and impede the proliferation of these cells 3 . Three genes encoding EF1α are present in the fission yeast genome. Previously, we determined that antibodies against Saccharomyces cerevisiae elongation factor proteins EF1α, EF2, and EF3 were effective in detecting their homologous forms in S. pombe . We found that deletions of EF1α‐a and EF1α‐c resulted in a slight decrease in total EF1α protein levels by utilizing Western blot analysis. These deletions produced no noticeable defect in cell growth on plates or in liquid culture. Continuing these studies, we plan to knock out EF1α‐b and analyze protein production. We will conduct phalloidin staining on our deletion mutants to determine if gene deletion has any visible effect on the polymerization of actin. Furthermore, we will knock out all three genes encoding EF1α in a single strain to assess its essentiality.