(R)-β-lysine-modified elongation factor P functions in translation elongation

TY - JOUR

T1 - (R)-β-lysine-modified elongation factor P functions in translation elongation

AU - Bullwinkle, Tammy J

AU - Zou, S Betty

AU - Rajkovic, Andrei

AU - Hersch, Steven J

AU - Elgamal, Sara

AU - Robinson, Nathaniel

AU - Smil, David

AU - Bolshan, Yuri

AU - Navarre, William Wiley

AU - Ibba, Michael

PY - 2013/2/8

Y1 - 2013/2/8

N2 - Post-translational modification of bacterial elongation factor P (EF-P) with (R)-β-lysine at a conserved lysine residue activates the protein in vivo and increases puromycin reactivity of the ribosome in vitro. The additional hydroxylation of EF-P at the same lysine residue by the YfcM protein has also recently been described. The roles of modified and unmodified EF-P during different steps in translation, and how this correlates to its physiological role in the cell, have recently been linked to the synthesis of polyproline stretches in proteins. Polysome analysis indicated that EF-P functions in translation elongation, rather than initiation as proposed previously. This was further supported by the inability of EF-P to enhance the rate of formation of fMet-Lys or fMet-Phe, indicating that the role of EF-P is not to specifically stimulate formation of the first peptide bond. Investigation of hydroxyl-(β)-lysyl-EF-P showed 30% increased puromycin reactivity but no differences in dipeptide synthesis rates when compared with the β-lysylated form. Unlike disruption of the other genes required for EF-P modification, deletion oyfcM had no phenotypic consequences in Salmonella. Taken together, our findings indicate that EF-P functions in translation elongation, a role critically dependent on post-translational β-lysylation but not hydroxylation.

AB - Post-translational modification of bacterial elongation factor P (EF-P) with (R)-β-lysine at a conserved lysine residue activates the protein in vivo and increases puromycin reactivity of the ribosome in vitro. The additional hydroxylation of EF-P at the same lysine residue by the YfcM protein has also recently been described. The roles of modified and unmodified EF-P during different steps in translation, and how this correlates to its physiological role in the cell, have recently been linked to the synthesis of polyproline stretches in proteins. Polysome analysis indicated that EF-P functions in translation elongation, rather than initiation as proposed previously. This was further supported by the inability of EF-P to enhance the rate of formation of fMet-Lys or fMet-Phe, indicating that the role of EF-P is not to specifically stimulate formation of the first peptide bond. Investigation of hydroxyl-(β)-lysyl-EF-P showed 30% increased puromycin reactivity but no differences in dipeptide synthesis rates when compared with the β-lysylated form. Unlike disruption of the other genes required for EF-P modification, deletion oyfcM had no phenotypic consequences in Salmonella. Taken together, our findings indicate that EF-P functions in translation elongation, a role critically dependent on post-translational β-lysylation but not hydroxylation.

KW - Bacterial Proteins

KW - Escherichia coli

KW - Hydroxylation

KW - Lysine

KW - Mixed Function Oxygenases

KW - Peptide Chain Elongation, Translational

KW - Peptide Elongation Factors

KW - Protein Processing, Post-Translational

KW - Salmonella enterica

U2 - 10.1074/jbc.M112.438879

DO - 10.1074/jbc.M112.438879

M3 - Journal article

C2 - 23277358

SN - 0021-9258

VL - 288

SP - 4416

EP - 4423

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 6

ER -