Identification and evaluation of regulatory factors modulating the expression of LEE4 in enterohemorrhagic Escherichia coli O157:H7

dc.contributor.author Mesterhazy, Karla
dc.contributor.department Veterinary Microbiology and Preventive Medicine
dc.date 2018-08-22T16:40:12.000
dc.date.accessioned 2020-06-30T08:00:45Z
dc.date.available 2020-06-30T08:00:45Z
dc.date.copyright Sun Jan 01 00:00:00 UTC 2006
dc.date.issued 2006-01-01
dc.description.abstract <p>The locus of enterocyte effacement (LEE) encoded operons (LEE1 - LEE5) enable enterohemorrhagic Escherichia coli (EHEC) O157:H7 to produce attaching and effacing lesions on intestinal cells (A/E). We have previously reported that hha-deleted EHEC O157:H7 up-regulated LEE4 through increased expression of ler, the regulatory gene encoded by the LEE1 operon. In the present study, we demonstrate that a transpositional insertion in clpXP abrogated the increased expression of the LEE4 in the hha-deleted strain. The loss of LEE4 up-regulation in an EHEC O157:H7 [Delta]hha, [Delta]clpXP mutant was restored by transposon insertions immediately upstream of grlR of the grlR-grlA operon, suggesting that grIR represses the expression of LEE4 in the absence of Hha and ClpXP. RT-PCR analysis revealed that grlR transcription increased by about 100-fold in strains lacking hha but remained the same in the presence or absence of clpXP thus indicating that hha acts as a negative regulator of grlR transcription. RT-PCR data also showed a 10-fold reduction in LEE4 transcription for EHEC O157:H7 [Delta]clpXP thus suggesting that the absence of clpXP resulted in an increased accumulation of GrlR and presumably Hha thereby causing an additional reduction in LEE4 transcription compared to that observed for LEE4 in EHEC O157:H7. Western blot analysis of the cell-free extracts of EHEC O157:H7 [Delta]clpXP revealed increased accumulation of Hha. Altogether, the results described in this report indicate that ClpXP is important for maintaining a critical intracellular pool of Hha, which in turn modulates transcription of grlR-grlA so that a basal level of transcription could continue through LEE4 and the other LEE operons.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/rtd/17535/
dc.identifier.articleid 18535
dc.identifier.contextkey 12371054
dc.identifier.doi https://doi.org/10.31274/rtd-180813-8317
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath rtd/17535
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/71359
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/rtd/17535/Mesterhazy_ISU_2006_M47.pdf|||Fri Jan 14 21:25:07 UTC 2022
dc.subject.keywords Veterinary Microbiology
dc.title Identification and evaluation of regulatory factors modulating the expression of LEE4 in enterohemorrhagic Escherichia coli O157:H7
dc.type article
dc.type.genre thesis
dspace.entity.type Publication
relation.isOrgUnitOfPublication 16f8e472-b1cd-4d8f-b016-09e96dbc4d83
thesis.degree.discipline Veterinary Microbiology
thesis.degree.level thesis
thesis.degree.name Master of Science
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