H-NST induces LEE expression and the formation of attaching and effacing lesions in enterohemorrhagic Escherichia coli.
Enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli are important causes of morbidity and mortality worldwide. These enteric pathogens contain a type III secretion system (T3SS) responsible for the attaching and effacing (A/E) lesion phenotype. The T3SS is encoded by the locus of enterocyt...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3897749?pdf=render |
| _version_ | 1818043255094247424 |
|---|---|
| author | Jonathan A Levine Anne-Marie Hansen Jane M Michalski Tracy H Hazen David A Rasko James B Kaper |
| author_facet | Jonathan A Levine Anne-Marie Hansen Jane M Michalski Tracy H Hazen David A Rasko James B Kaper |
| author_sort | Jonathan A Levine |
| collection | DOAJ |
| description | Enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli are important causes of morbidity and mortality worldwide. These enteric pathogens contain a type III secretion system (T3SS) responsible for the attaching and effacing (A/E) lesion phenotype. The T3SS is encoded by the locus of enterocyte effacement (LEE) pathogenicity island. The H-NS-mediated repression of LEE expression is counteracted by Ler, the major activator of virulence gene expression in A/E pathogens. A regulator present in EPEC, H-NST, positively affects expression of H-NS regulon members in E. coli K-12, although the effect of H-NST on LEE expression and virulence of A/E pathogens has yet-to-be determined.We examine the effect of H-NST on LEE expression and A/E lesion formation on intestinal epithelial cells. We find that H-NST positively affects the levels of LEE-encoded proteins independently of ler and induces A/E lesion formation. We demonstrate H-NST binding to regulatory regions of LEE1 and LEE3, the first report of DNA-binding by H-NST. We characterize H-NST mutants substituted at conserved residues including Ala16 and residues Arg60 and Arg63, which are part of a potential DNA-binding domain. The single mutants A16V, A16L, R60Q and the double mutant R60Q/R63Q exhibit a decreased effect on LEE expression and A/E lesion formation. DNA mobility shift assays reveal that these residues are important for H-NST to bind regulatory LEE DNA targets. H-NST positively affects Ler binding to LEE DNA in the presence of H-NS, and thereby potentially helps Ler displace H-NS bound to DNA.H-NST induces LEE expression and A/E lesion formation likely by counteracting H-NS-mediated repression. We demonstrate that H-NST binds to DNA and identify arginine residues that are functionally important for DNA-binding. Our study suggests that H-NST provides an additional means for A/E pathogens to alleviate repression of virulence gene expression by H-NS to promote virulence capabilities. |
| first_indexed | 2024-12-10T08:59:18Z |
| format | Article |
| id | doaj.art-2cada9974ef84a86ab2cb958e2ddf072 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-10T08:59:18Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-2cada9974ef84a86ab2cb958e2ddf0722022-12-22T01:55:20ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0191e8661810.1371/journal.pone.0086618H-NST induces LEE expression and the formation of attaching and effacing lesions in enterohemorrhagic Escherichia coli.Jonathan A LevineAnne-Marie HansenJane M MichalskiTracy H HazenDavid A RaskoJames B KaperEnteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli are important causes of morbidity and mortality worldwide. These enteric pathogens contain a type III secretion system (T3SS) responsible for the attaching and effacing (A/E) lesion phenotype. The T3SS is encoded by the locus of enterocyte effacement (LEE) pathogenicity island. The H-NS-mediated repression of LEE expression is counteracted by Ler, the major activator of virulence gene expression in A/E pathogens. A regulator present in EPEC, H-NST, positively affects expression of H-NS regulon members in E. coli K-12, although the effect of H-NST on LEE expression and virulence of A/E pathogens has yet-to-be determined.We examine the effect of H-NST on LEE expression and A/E lesion formation on intestinal epithelial cells. We find that H-NST positively affects the levels of LEE-encoded proteins independently of ler and induces A/E lesion formation. We demonstrate H-NST binding to regulatory regions of LEE1 and LEE3, the first report of DNA-binding by H-NST. We characterize H-NST mutants substituted at conserved residues including Ala16 and residues Arg60 and Arg63, which are part of a potential DNA-binding domain. The single mutants A16V, A16L, R60Q and the double mutant R60Q/R63Q exhibit a decreased effect on LEE expression and A/E lesion formation. DNA mobility shift assays reveal that these residues are important for H-NST to bind regulatory LEE DNA targets. H-NST positively affects Ler binding to LEE DNA in the presence of H-NS, and thereby potentially helps Ler displace H-NS bound to DNA.H-NST induces LEE expression and A/E lesion formation likely by counteracting H-NS-mediated repression. We demonstrate that H-NST binds to DNA and identify arginine residues that are functionally important for DNA-binding. Our study suggests that H-NST provides an additional means for A/E pathogens to alleviate repression of virulence gene expression by H-NS to promote virulence capabilities.http://europepmc.org/articles/PMC3897749?pdf=render |
| spellingShingle | Jonathan A Levine Anne-Marie Hansen Jane M Michalski Tracy H Hazen David A Rasko James B Kaper H-NST induces LEE expression and the formation of attaching and effacing lesions in enterohemorrhagic Escherichia coli. PLoS ONE |
| title | H-NST induces LEE expression and the formation of attaching and effacing lesions in enterohemorrhagic Escherichia coli. |
| title_full | H-NST induces LEE expression and the formation of attaching and effacing lesions in enterohemorrhagic Escherichia coli. |
| title_fullStr | H-NST induces LEE expression and the formation of attaching and effacing lesions in enterohemorrhagic Escherichia coli. |
| title_full_unstemmed | H-NST induces LEE expression and the formation of attaching and effacing lesions in enterohemorrhagic Escherichia coli. |
| title_short | H-NST induces LEE expression and the formation of attaching and effacing lesions in enterohemorrhagic Escherichia coli. |
| title_sort | h nst induces lee expression and the formation of attaching and effacing lesions in enterohemorrhagic escherichia coli |
| url | http://europepmc.org/articles/PMC3897749?pdf=render |
| work_keys_str_mv | AT jonathanalevine hnstinducesleeexpressionandtheformationofattachingandeffacinglesionsinenterohemorrhagicescherichiacoli AT annemariehansen hnstinducesleeexpressionandtheformationofattachingandeffacinglesionsinenterohemorrhagicescherichiacoli AT janemmichalski hnstinducesleeexpressionandtheformationofattachingandeffacinglesionsinenterohemorrhagicescherichiacoli AT tracyhhazen hnstinducesleeexpressionandtheformationofattachingandeffacinglesionsinenterohemorrhagicescherichiacoli AT davidarasko hnstinducesleeexpressionandtheformationofattachingandeffacinglesionsinenterohemorrhagicescherichiacoli AT jamesbkaper hnstinducesleeexpressionandtheformationofattachingandeffacinglesionsinenterohemorrhagicescherichiacoli |