Study of acyltransferases required for the synthesis of membrane phospholipids in Gram-positive bacteria: an attractive target for antibacterial drug discovery

• Grant number: 14/13411-1
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): November 01, 2014
• Effective date (End): October 31, 2017
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Frederico José Gueiros Filho
• Grantee: Diego Emiliano Sastre
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The biosynthesis of membrane lipids is an essential pathway for virtually all bacteria. Although the spectrum of phospholipid headgroup structures produced by bacteria is large, the key precursor of these molecules is phosphatidic acid (PtdOH). Glycerol-3-phosphate derived from the glycolysis is the universal source for the glycerol backbone of PtdOH. There are two distinct pathways responsible for the acylation of glycerol-3-phosphate. The PlsB/PlsC acyltransferases were discovered in Escherichia coli, and homologs are present in many eukaryotes. This protein family primarily uses acyl-acyl carrier protein (ACP) endproducts of fatty acid synthesis as acyl donors, but may also use acyl-CoA derived from exogenous fatty acids. The second pathway, PlsX/PlsY/PlsC, is more widely distributed in bacteria. The PlsY acyltransferase utilizes as unique acyl donor, acyl-phosphate, which is produced from acyl-ACP by the key enzyme PlsX. The acylation of the 2-position is carried out by members of the PlsC protein family. Despite its potential importance for the development of novel antibiotics, little is known about the underlying signaling mechanisms that allow bacteria to control their membrane lipid composition within narrow limits. The regulation of PlsX activity remains a major unanswered question in bacterial biochemistry and physiology. Understanding this enzyme is particularly relevant in light of its proposed role in the coordination of fatty acid and phospholipid synthesis in the major human pathogen Gram-positive bacteria. Also, genetic regulation of the PlsX/PlsY/PlsC pathway as the sole route to membrane phospholipids remains an important avenue to explore. These essential enzymes are unique to bacteria and thus represent a new attractive target for the development of novel antibacterial agents, as well as, the global transcriptional regulator FapR controlling the PlsX and PlsC expression. In this study, we proposed to study the mechanisms of regulation of acyltransferase activity PlsX and to determine the possible association of PlsX with membranes and/or the division of machinery. Also, we will try to identify putative inhibitors of the key enzymes that regulate lipid metabolism in Gram-positive bacteria (FapR and PlsX) to combat the growing problem of pathogens resistance to current antibiotics. (AU)

Scientific publications

(6)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MODENEZ, IAGO A.; SASTRE, DIEGO E.; MORAES, FERNANDO C.; MARQUES NETTO, CATERINA G. C.. Influence of Glutaraldehyde Cross-Linking Modes on the Recyclability of Immobilized Lipase B from Candida antarctica for Transesterification of Soy Bean Oil. Molecules, v. 23, n. 9, SEP 2018. (16/01622-3, 14/13411-1)

SASTRE, DIEGO E.; SANTOS, LEIDAIANY P.; KAGOHARA, EDNA; ANDRADE, LEANDRO H.. Draft Whole-Genome Sequence of Psychrotrophic Arthrobacter sp. Strain 7749, Isolated from Antarctic Marine Sediments with Applications in Enantioselective Alcohol Oxidation. MICROBIOLOGY RESOURCE ANNOUNCEMENTS, v. 5, n. 43, OCT 2017. (14/13411-1, 17/02854-8)

SASTRE, DIEGO EMILIANO; BISSON-FILHO, ALEXANDRE; DE MENDOZA, DIEGO; GUEIROS-FILHO, FREDERICO J.. Revisiting the cell biology of the acyl-ACP: phosphate transacylase PlsX suggests that the phospholipid synthesis and cell division machineries are not coupled in Bacillus subtilis. Molecular Microbiology, v. 100, n. 4, p. 621-634, MAY 2016. (14/13411-1)

PULSCHEN, ANDRE A.; SASTRE, DIEGO E.; MACHINANDIARENA, FEDERICO; ASIS, AGOSTINA CROTTA; ALBANESI, DANIELA; DE MENDOZA, DIEGO; GUEIROS-FILHO, FREDERICO J.. The stringent response plays a key role in Bacillus subtilis survival of fatty acid starvation. Molecular Microbiology, v. 103, n. 4, p. 698-712, FEB 2017. (14/26528-4, 14/13411-1)

SASTRE, DIEGO E.; BASSO, LUIS G. M.; TRASTOY, BEATRIZ; CIFUENTE, JAVIER O.; CONTRERAS, XABIER; GUEIROS-FILHO, FREDERICO; DE MENDOZA, DIEGO; NAVARRO, MARCOS V. A. S.; GUERIN, MARCELO E.. Membrane fluidity adjusts the insertion of the transacylase PlsX to regulate phospholipid biosynthesis in Gram-positive bacteria. Journal of Biological Chemistry, v. 295, n. 7, p. 2136-2147, FEB 14 2020. (16/05203-5, 15/21583-0, 14/00206-0, 14/13411-1)

SASTRE, DIEGO E.; PULSCHEN, ANDRE A.; BASSO, LUIS G. M.; PARIENTE, JHONATHAN S. BENITES; MARQUES NETTO, CATERINA G. C.; MACHINANDIARENA, FEDERICO; ALBANESI, DANIELA; NAVARRO, MARCOS V. A. S.; DE MENDOZA, DIEGO; GUEIROS-FILHO, FREDERICO J.. The phosphatidic acid pathway enzyme PlsX plays both catalytic and channeling roles in bacterial phospholipid synthesis. Journal of Biological Chemistry, v. 295, n. 7, p. 2148-2159, FEB 14 2020. (16/05203-5, 15/21583-0, 14/00206-0, 14/13411-1)