Truncation of the Essential Escherichia coli K-12 Inner Membrane Protein, YejM, Results in Temperature-Sensitive Growth: Temperature-Sensitivity and Essentiality are Suppressed by Overexpression of the AcpT Phosphopantetheinyl Transferase

Strain LH530 a mutant of Escherichia coli K-12 was reported by others to show increased outer membrane permeability, temperature-sensitive growth and reduced synthesis of lipid A. The unmapped mutant gene was found to be suppressed by high copy number plasmids carrying the wild type acpT gene which encodes a protein that catalyzes a post-translational protein modification, the attachment of 4' phosphopantetheine. We mapped the strain LH530 mutation to a gene of unknown function, yejM, known to encode an inner membrane protein. The mutation is a yejM nonsense mutation that produces a truncated protein lacking the predicted periplasmic domain. Reconstruction of the mutation gave a strain having the same phenotypes as LH530. In contrast to the nonsense mutants, deletion of the entire yejM gene was lethal. Suppression by AcpT overexpression of the yejM nonsense mutants encoding the truncated proteins was specific to AcpT. Moreover, AcpT overexpression also suppressed the lethality due to deletion of the entire yejM gene and this suppression also did not require that AcpT be enzymatically active. The mechanism whereby overexpression of a specific cytosolic protein bypasses the essentiality of an inner membrane protein is unknown.

Key Words: acpT, inner membrane protein, phosphopantetheinyl transferase, temperature-sensitive growth, yejM

Online ISS 1091-6490
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