The inner membrane protein YhdP modulates the rate of anterograde phospholipid flow in <i>Escherichia coli</i>

Abstract

The outer membrane (OM) of Gram-negative bacteria is a selective permeability barrier that allows uptake of nutrients while simultaneously protecting the cell from harmful compounds. The basic pathways and molecular machinery responsible for transporting lipopolysaccharides (LPS), lipoproteins, and β-barrel proteins to the OM have been identified, but very little is known about phospholipid (PL) transport. To identify genes capable of affecting PL transport, we screened for genetic interactions with <i>mlaA</i>*, a mutant in which anterograde PL transport causes the inner membrane (IM) to shrink and eventually rupture; characterization of <i>mlaA</i>*-mediated lysis suggested that PL transport can occur via a high-flux diffusive flow mechanism. We found that YhdP, an IM protein involved in maintaining the OM permeability barrier, modulates the rate of PL transport during <i>mlaA</i>*-mediated lysis. Deletion of <i>yhdP</i> from <i>mlaA</i>* reduced the rate of IM transport to the OM by 50%, slowing shrinkage of the IM and delaying lysis. As a result, the weakened OM of ∆<i>yhdP</i> cells was further compromised and ruptured before the IM during <i>mlaA</i>*-mediated death. These findings demonstrate the existence of a high-flux diffusive pathway for PL flow in <i>Escherichia coli</i> that is modulated by YhdP.