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Journal of Bacteriology, January 2007, p. 244-253, Vol. 189, No. 1
0021-9193/07/$08.00+0     doi:10.1128/JB.01126-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Characterization of lptA and lptB, Two Essential Genes Implicated in Lipopolysaccharide Transport to the Outer Membrane of Escherichia coli{triangledown}

Paola Sperandeo,1 Rachele Cescutti,2 Riccardo Villa,2 Cristiano Di Benedetto,3 Daniela Candia,3 Gianni Dehò,1 and Alessandra Polissi2*

Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, Milan, Italy,1 Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Milan, Italy,2 Dipartimento di Biologia, Università degli Studi di Milano, Milan, Italy3

Received 27 July 2006/ Accepted 11 October 2006

The outer membrane (OM) of gram-negative bacteria is an asymmetric lipid bilayer that protects the cell from toxic molecules. Lipopolysaccharide (LPS) is an essential component of the OM in most gram-negative bacteria, and its structure and biosynthesis are well known. Nevertheless, the mechanisms of transport and assembly of this molecule in the OM are poorly understood. To date, the only proteins implicated in LPS transport are MsbA, responsible for LPS flipping across the inner membrane, and the Imp/RlpB complex, involved in LPS targeting to the OM. Here, we present evidence that two Escherichia coli essential genes, yhbN and yhbG, now renamed lptA and lptB, respectively, participate in LPS biogenesis. We show that mutants depleted of LptA and/or LptB not only produce an anomalous LPS form, but also are defective in LPS transport to the OM and accumulate de novo-synthesized LPS in a novel membrane fraction of intermediate density between the inner membrane (IM) and the OM. In addition, we show that LptA is located in the periplasm and that expression of the lptA-lptB operon is controlled by the extracytoplasmic {sigma} factor RpoE. Based on these data, we propose that LptA and LptB are implicated in the transport of LPS from the IM to the OM of E. coli.

* Corresponding author. Mailing address: Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca, Piazza della Scienza 220126, Milano, Italy. Phone: 39-02-64483431. Fax: 39-02-6448-3450. E-mail: alessandra.polissi{at}unimib.it.

{triangledown} Published ahead of print on 20 October 2006.

Journal of Bacteriology, January 2007, p. 244-253, Vol. 189, No. 1
0021-9193/07/$08.00+0     doi:10.1128/JB.01126-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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