Unexpected biochemistry determines endotoxin structure in two enteric gram-negatives
| dc.contributor.advisor | Trent, Michael Stephen | en |
| dc.contributor.committeeMember | Davies, Bryan W | en |
| dc.contributor.committeeMember | Ehrlich, Lauren | en |
| dc.contributor.committeeMember | Maynard, Jennifer | en |
| dc.contributor.committeeMember | Payne, Shelley | en |
| dc.creator | Di Pierro, Erica Jacqueline | en |
| dc.date.accessioned | 2015-08-25T17:12:45Z | en |
| dc.date.issued | 2015-05 | en |
| dc.date.submitted | May 2015 | en |
| dc.date.updated | 2015-08-25T17:12:45Z | en |
| dc.description.abstract | Most gram-negative organisms require lipopolysaccharide and its membrane anchor, lipid A, for growth and survival. Also known as endotoxin, lipid A is synthesized via a nine-step enzymatic process, culminating in a conserved hexa-acylated, bis-phosphorylated disaccharide of glucosamine. This framework is often altered by condition- or species-specific lipid A modifications, which change the biochemical properties of the molecule in response to and to defend against environmental stress signals. Here, we expound on two stories in different gram-negative organisms, both involving novel or unanticipated biochemistry that impacts lipid A structure. First, the missing acyltransferase in the Epsilonproteobacterium Helicobacter pylori lipid A biosynthesis pathway is identified. This enzyme transfers a secondary acyl chain to the 3'-linked primary acyl chain of lipid A like E. coli LpxM, but shares almost no sequence similarity with the E. coli acyltransferase. It is reannotated as LpxJ and demonstrated to possess an unprecedented ability to act before the 2'-secondary acyltransferase, LpxL, as well as the 3-deoxy-D-manno-octulosonic acid transferase, KdtA. LpxJ is one member of a large class of acyltransferases found in a diverse range of organisms that lack an E. coli LpxM homolog, suggesting that LpxJ participates in lipid A biosynthesis in place of an LpxM homolog. The second story focuses on regulation of modifications to endotoxin structure that occur after the conserved biosynthesis pathway. E. coli pmrD is shown to be required for PmrAB-dependent lipid A modifications in conditions that exclusively activate PhoPQ; this result proves that PmrD connects PhoPQ and PmrAB despite previous reports that it is an inactive connector in this organism. Further, RNA sequencing and polymyxin B survival assays solidify the role of E. coli pmrD in influencing expression of pmrA and its target genes and promoting survival during exposure to cationic antimicrobial peptides. Notably, the presence of an unknown factor or system capable of activating pmrD to promote lipid A modification in the absence of the PhoPQ system is also revealed. In all, the findings presented here expand our understanding of alternative approaches to lipid A biosynthesis and the complex systems that regulate modifications of this dynamic molecule. | en |
| dc.description.department | Cellular and Molecular Biology | en |
| dc.format.mimetype | application/pdf | en |
| dc.identifier.uri | http://hdl.handle.net/2152/30448 | en |
| dc.language.iso | en | en |
| dc.subject | Lipid A | en |
| dc.subject | Helicobacter pylori | en |
| dc.subject | Escherichia coli | en |
| dc.subject | Acyltransferase | en |
| dc.subject | Lipid A biosynthesis | en |
| dc.subject | Lipid A modifications | en |
| dc.subject | Cationic antimicrobial peptide resistance | en |
| dc.title | Unexpected biochemistry determines endotoxin structure in two enteric gram-negatives | en |
| dc.type | Thesis | en |
| thesis.degree.department | Cellular and Molecular Biology | en |
| thesis.degree.discipline | Cellular and Molecular Biology | en |
| thesis.degree.grantor | The University of Texas at Austin | en |
| thesis.degree.level | Doctoral | en |
| thesis.degree.name | Doctor of Philosophy | en |