title: Inhibition of cell division induced by external guide sequences (EGS Technology) targeting ftsZ.
abstract: EGS (external guide sequence) technology is a promising approach to designing new antibiotics. EGSs are short antisense oligoribonucleotides that induce RNase P-mediated cleavage of a target RNA by forming a precursor tRNA-like complex. The ftsZ mRNA secondary structure was modeled and EGSs complementary to two regions with high probability of being suitable targets were designed. In vitro reactions showed that EGSs targeting these regions bound ftsZ mRNA and elicited RNase P-mediated cleavage of ftsZ mRNA. A recombinant plasmid, pEGSb1, coding for an EGS that targets region "b" under the control of the T7 promoter was generated. Upon introduction of this plasmid into Escherichia coli BL21(DE3)(pLysS) the transformant strain formed filaments when expression of the EGS was induced. Concomitantly, E. coli harboring pEGSb1 showed a modest but significant inhibition of growth when synthesis of the EGSb1 was induced. Our results indicate that EGS technology could be a viable strategy to generate new antimicrobials targeting ftsZ.
authors.affiliation: Fundación Instituto Leloir-Instituto de Investigaciones Bioquímicas Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas - Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina.
authors.forename: Carol Davies
authors.initials: CD
authors.lastname: Sala
authors.forename: Alfonso J C
authors.initials: AJ
authors.lastname: Soler-Bistué
authors.forename: Leeann
authors.initials: L
authors.lastname: Korprapun
authors.forename: Angeles
authors.initials: A
authors.lastname: Zorreguieta
authors.forename: Marcelo E
authors.initials: ME
authors.lastname: Tolmasky
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D015195:Drug Design; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D020320:Oligoribonucleotides, Antisense / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D059367:RNA Cleavage / Q000187:drug effects*; D043262:Ribonuclease P / Q000378:metabolism*; D013728:Terminator Regions, Genetic / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; C071525:FtsZ protein, Bacteria; D020320:Oligoribonucleotides, Antisense; D043262:Ribonuclease P