Elongation factor 1beta is an actin-binding protein.
A 17 kDa polypeptide found in association with actin in cellular extracts of Dictyostelium discoideum was identified as a proteolytic fragment of eEF1beta. Antibody elicited against the 17 kDa protein reacted with a single 29 kDa polypeptide in Dictyostelium, indicating that the 17 kDa peptide arises from degradation of a larger precursor. The cDNA isolated from a Dictyostelium library using this antibody as a probe encodes Dictyostelium elongation factor 1beta. Amino acid degradation of the 17 kDa protein fragment confirmed the identity of the protein as eEF1beta. Direct interaction of eEF1beta with actin in vitro was further demonstrated in mixtures of actin with the 17 kDa protein fragment of Dictyostelium eEF1beta, recombinant preparations of Dictyostelium eEF1beta expressed in Escherichia coli, and the intact eEF1betagamma complex purified from wheat germ. Localization of eEF1beta in Dictyostelium by immunofluorescence microscopy reveals both diffuse cytoplasmic staining, and some concentration in the cortical and hyaline cytoplasm. The results support the existence of physical and functional interactions of the translation apparatus with the cytoskeleton, and suggest that eEF1beta may function in a dual role both to promote the elongation phase of protein synthesis, and to interact with cytoplasmic actin.
Furukawa
R
R
Department of Cellular Biology, University of Georgia, 724 Biological Science Building, Athens, GA 30602, USA.
Jinks
T M
TM
Tishgarten
T
T
Mazzawi
M
M
Morris
D R
DR
Fechheimer
M
M
D000199:Actins / Q000737:chemistry / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000737:chemistry; D004023:Dictyostelium / Q000737:chemistry* / Q000235:genetics; D015723:Gene Library; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D020648:Peptide Elongation Factor 1 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D015183:Restriction Mapping; D016415:Sequence Alignment
D000199:Actins; D000906:Antibodies; D008840:Microfilament Proteins; D020648:Peptide Elongation Factor 1