title: A putative lytic transglycosylase tightly regulated and critical for the EHEC type three secretion.
abstract: Open reading frame l0045 in the pathogenic island of enterohemorrhagic Escherichia coli O157:H7 has been predicted to encode a lytic transglycosylase that is homologous to two different gene products encoded by the same bacteria at loci away from the island. To deduce the necessity of the presence in the island, we created an l0045-deleted strain of EHEC and observed that both the level of cytosolic EspA and that of the other type III secreted proteins in the media were affected. In a complementation assay, a low level-expressing L0045 appeared to recover efficiently the type III secretion (TTS). On the other hand, when l0045 was driven to express robustly, the intracellular levels of representative TTS proteins were severely suppressed. This suppression is apparently caused by the protein of L0045 per se since introducing an early translational termination codon abolished the suppression. Intriguingly, the authentic L0045 was hardly detected in all lysates of EHEC differently prepared while the same construct was expectedly expressed in the K-12 strain. A unique network must exist in EHEC to tightly regulate the presence of L0045, and we found that a LEE regulator (GrlA) is critically involved in this regulation.
authors.affiliation: Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Yen-Chi
authors.initials: YC
authors.lastname: Yu
authors.forename: Ching-Nan
authors.initials: CN
authors.lastname: Lin
authors.forename: Shao-Hung
authors.initials: SH
authors.lastname: Wang
authors.forename: Swee-Chuan
authors.initials: SC
authors.lastname: Ng
authors.forename: Wensi S
authors.initials: WS
authors.lastname: Hu
authors.forename: Wan-Jr
authors.initials: WJ
authors.lastname: Syu
mesh_terms: D017931:DNA Primers / Q000235:genetics; D019453:Escherichia coli O157 / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000378:metabolism; D005816:Genetic Complementation Test; D044404:Genomic Islands / Q000235:genetics*; D016695:Glycosyltransferases / Q000235:genetics*; D015151:Immunoblotting; D016366:Open Reading Frames / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055571:Secretory Pathway / Q000235:genetics / Q000502:physiology*; D015534:Trans-Activators / Q000378:metabolism
chemical_list: D017931:DNA Primers; D029968:Escherichia coli Proteins; C102051:EspA protein, E coli; C513430:GrlA protein, E coli; D015534:Trans-Activators; D016695:Glycosyltransferases