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Molecular cloning and characterization of the Arabidopsis thaliana α‐subunit of elongation factor 1B
Author(s) -
Héricourt François,
Jupin Isabelle
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01694-4
Subject(s) - biology , elongation factor , protein subunit , arabidopsis , complementary dna , arabidopsis thaliana , open reading frame , homology (biology) , genetics , gene , guanine nucleotide exchange factor , microbiology and biotechnology , mutant , peptide sequence , rna , ribosome , gtpase
Using a PCR‐based approach, we have isolated two Arabidopsis thaliana cDNA clones (α1 and α2) encoding the α‐subunit of translation elongation factor 1B (eEF1Bα). They encode open reading frames of 228 and 224 amino acids respectively, with extensive homology to eEF1Bα subunits from different organisms, particularly in the C‐terminal half of the protein. They both lack a conserved phosphorylation site that has been implicated in regulating nucleotide exchange activity. Using a plasmid shuffling experiment, we demonstrated that both α1 and α2 clones are able to complement a mutant yeast strain deficient for the eEF1Bα subunit. This provides evidence that Arabidopsis encodes at least two functional isoforms of this subunit, termed eEF1Bα1 and eEF1Bα2. A third cDNA clone was isolated that appeared to result from an alternative splicing event of the eEF1Bα1 gene.