Eukaryotic initiation factor 5A dephosphorylation is required for translational arrest in stationary phase cells.
The protein known as eIF5A (eukaryotic initiation factor 5A) has an elusive role in translation. It has a unique and essential hypusine modification at a conserved lysine residue in most eukaryotes. In addition, this protein is modified by phosphorylation with unknown functions. In the present study we show that a phosphorylated state of eIF5A predominates in exponentially growing Trypanosoma cruzi cells, and extensive dephosphorylation occurs in cells in stationary phase. Phosphorylation occurs mainly at Ser(2), as shown in yeast eIF5A. In addition, a novel phosphorylation site was identified at Tyr(21). In exponential cells, T. cruzi eIF5A is partially associated with polysomes, compatible with a proposed function as an elongation factor, and becomes relatively enriched in polysomal fractions in stationary phase. Overexpression of the wild-type eIF5A, or eIF5A with Ser(2) replaced by an aspartate residue, but not by alanine, increases the rate of cell proliferation and protein synthesis. However, the presence of an aspartate residue instead of Ser(2) is toxic for cells reaching the stationary phase, which show a less-pronounced protein synthesis arrest and a decreased amount of eIF5A in dense fractions of sucrose gradients. We conclude that eIF5A phosphorylation and dephosphorylation cycles regulate translation according to the growth conditions.
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Rua Pedro de Toledo 669 L6A, São Paulo, S.P. 04039-032, Brazil.
Janete
J
Chung
Antonio A
AA
Rocha
Renata R
RR
Tonelli
Beatriz A
BA
Castilho
Sergio
S
Schenkman
D000409:Alanine / Q000378:metabolism; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001224:Aspartic Acid / Q000378:metabolism; D008969:Molecular Sequence Data; D010448:Peptide Initiation Factors / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000166:cytology / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism; D000097574:Eukaryotic Translation Initiation Factor 5A
D000409:Alanine; D001224:Aspartic Acid; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D012694:Serine; D014443:Tyrosine; D000097574:Eukaryotic Translation Initiation Factor 5A