Molecular and structural basis of inner core lipopolysaccharide alterations in Escherichia coli: incorporation of glucuronic acid and phosphoethanolamine in the heptose region.

Abstract

It is well established that lipopolysaccharide (LPS) often carries nonstoichiometric substitutions in lipid A and in the inner core. In this work, the molecular basis of inner core alterations and their physiological significance are addressed. A new inner core modification of LPS is described, which arises due to the addition of glucuronic acid on the third heptose with a concomitant loss of phosphate on the second heptose. This was shown by chemical and structural analyses. Furthermore, the gene whose product is responsible for the addition of this sugar was identified in all Escherichia coli core types and in Salmonella and was designated waaH. Its deduced amino acid sequence exhibits homology to glycosyltransferase family 2. The transcription of the waaH gene is positively regulated by the PhoB/R two-component system in a growth phase-dependent manner, which is coordinated with the transcription of the ugd gene explaining the genetic basis of this modification. Glucuronic acid modification was observed in E. coli B, K12, R2, and R4 core types and in Salmonella. We also show that the phosphoethanolamine (P-EtN) addition on heptose I in E. coli K12 requires the product of the ORF yijP, a new gene designated as eptC. Incorporation of P-EtN is also positively regulated by PhoB/R, although it can occur at a basal level without a requirement for any regulatory inducible systems. This P-EtN modification is essential for resistance to a variety of factors, which destabilize the outer membrane like the addition of SDS or challenge to sublethal concentrations of Zn(2+).

• Authors: Gracjana Klein, Sven Müller-Loennies, Buko Lindner, Natalia Kobylak, Helmut Brade, Satish Raina
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 288 Issue 12 Pg. 8111-8127 (Mar 22 2013) ISSN: 1083-351X [Electronic] United States
• PMID: 23372159 (Publication Type: Journal Article)

Chemical References

• Escherichia coli Proteins
• Ethanolamines
• Heptoses
• Lipopolysaccharides
• Membrane Proteins
• eptC protein, E coli
• phosphorylethanolamine
• Glucuronic Acid
• Glycosyltransferases
• WaaQ protein, E coli

Topics

• Carbohydrate Conformation
• Carbohydrate Sequence
• Cell Membrane (metabolism, physiology)
• Escherichia coli (genetics, metabolism)
• Escherichia coli Proteins (genetics, metabolism, physiology)
• Ethanolamines (metabolism)
• Gene Expression Regulation, Bacterial
• Glucuronic Acid (metabolism)
• Glycosyltransferases (genetics)
• Heptoses (metabolism)
• Lipopolysaccharides (chemistry, metabolism)
• Membrane Proteins (genetics, metabolism, physiology)
• Molecular Sequence Annotation
• Molecular Sequence Data
• Transcription, Genetic