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Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent
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| Zeitschriftentitel: | Microbiology |
|---|---|
| Personen und Körperschaften: | , , |
| In: | Microbiology, 151, 2005, 12, S. 3979-3987 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Microbiology Society
|
| Schlagwörter: |
| author_facet |
Perron-Savard, Philippe De Crescenzo, Gregory Moual, Hervé Le Perron-Savard, Philippe De Crescenzo, Gregory Moual, Hervé Le |
|---|---|
| author |
Perron-Savard, Philippe De Crescenzo, Gregory Moual, Hervé Le |
| spellingShingle |
Perron-Savard, Philippe De Crescenzo, Gregory Moual, Hervé Le Microbiology Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent Microbiology |
| author_sort |
perron-savard, philippe |
| spelling |
Perron-Savard, Philippe De Crescenzo, Gregory Moual, Hervé Le 1350-0872 1465-2080 Microbiology Society Microbiology http://dx.doi.org/10.1099/mic.0.28236-0 <jats:p>In<jats:italic>Salmonella enterica</jats:italic>, PhoP is the response regulator of the PhoP/PhoQ two-component regulatory system that controls the expression of various virulence factors in response to external Mg<jats:sup>2+</jats:sup>. Previous studies have shown that phosphorylation of a PhoP variant with a C-terminal His tag (PhoP<jats:sub>His</jats:sub>) enhances dimerization and binding to target DNA. Here, the effect of phosphorylation on the oligomerization and DNA binding properties of both wild-type PhoP (PhoP) and PhoP<jats:sub>His</jats:sub>are compared. Gel filtration chromatography showed that PhoP exists as a mixture of monomer and dimer regardless of its phosphorylation state. In contrast, unphosphorylated PhoP<jats:sub>His</jats:sub>was mostly monomeric, whereas PhoP<jats:sub>His</jats:sub>∼P existed as a mixture of monomer and dimer. By monitoring the tryptophan fluorescence of the proteins and the fluorescence of the probe 1-anilinonaphthalene-8-sulfonic acid bound to them, it was found that PhoP and PhoP<jats:sub>His</jats:sub>exhibited different spectral properties. The interaction between PhoP or PhoP<jats:sub>His</jats:sub>and the PhoP box of the<jats:italic>mgtA</jats:italic>promoter was monitored by surface plasmon resonance. Binding of PhoP to the PhoP box was barely influenced by phosphorylation. In contrast, phosphorylation of PhoP<jats:sub>His</jats:sub>clearly increased the interaction of PhoP<jats:sub>His</jats:sub>with target DNA. Altogether, these data show that a His tag at the C-terminus of PhoP affects its biochemical properties, most likely by affecting its conformation and/or its oligomerization state. More importantly, these results show that wild-type PhoP dimerization and interaction with target DNA are independent of phosphorylation, which is in contrast to the previously proposed model.</jats:p> Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent Microbiology |
| doi_str_mv |
10.1099/mic.0.28236-0 |
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Online Free |
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Biologie |
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Microbiology Society, 2005 |
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Microbiology Society, 2005 |
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1350-0872 1465-2080 |
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2005 |
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Microbiology Society |
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Microbiology |
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49 |
| title |
Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_unstemmed |
Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_full |
Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_fullStr |
Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_full_unstemmed |
Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_short |
Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_sort |
dimerization and dna binding of the salmonella enterica phop response regulator are phosphorylation independent |
| topic |
Microbiology |
| url |
http://dx.doi.org/10.1099/mic.0.28236-0 |
| publishDate |
2005 |
| physical |
3979-3987 |
| description |
<jats:p>In<jats:italic>Salmonella enterica</jats:italic>, PhoP is the response regulator of the PhoP/PhoQ two-component regulatory system that controls the expression of various virulence factors in response to external Mg<jats:sup>2+</jats:sup>. Previous studies have shown that phosphorylation of a PhoP variant with a C-terminal His tag (PhoP<jats:sub>His</jats:sub>) enhances dimerization and binding to target DNA. Here, the effect of phosphorylation on the oligomerization and DNA binding properties of both wild-type PhoP (PhoP) and PhoP<jats:sub>His</jats:sub>are compared. Gel filtration chromatography showed that PhoP exists as a mixture of monomer and dimer regardless of its phosphorylation state. In contrast, unphosphorylated PhoP<jats:sub>His</jats:sub>was mostly monomeric, whereas PhoP<jats:sub>His</jats:sub>∼P existed as a mixture of monomer and dimer. By monitoring the tryptophan fluorescence of the proteins and the fluorescence of the probe 1-anilinonaphthalene-8-sulfonic acid bound to them, it was found that PhoP and PhoP<jats:sub>His</jats:sub>exhibited different spectral properties. The interaction between PhoP or PhoP<jats:sub>His</jats:sub>and the PhoP box of the<jats:italic>mgtA</jats:italic>promoter was monitored by surface plasmon resonance. Binding of PhoP to the PhoP box was barely influenced by phosphorylation. In contrast, phosphorylation of PhoP<jats:sub>His</jats:sub>clearly increased the interaction of PhoP<jats:sub>His</jats:sub>with target DNA. Altogether, these data show that a His tag at the C-terminus of PhoP affects its biochemical properties, most likely by affecting its conformation and/or its oligomerization state. More importantly, these results show that wild-type PhoP dimerization and interaction with target DNA are independent of phosphorylation, which is in contrast to the previously proposed model.</jats:p> |
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| author | Perron-Savard, Philippe, De Crescenzo, Gregory, Moual, Hervé Le |
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| description | <jats:p>In<jats:italic>Salmonella enterica</jats:italic>, PhoP is the response regulator of the PhoP/PhoQ two-component regulatory system that controls the expression of various virulence factors in response to external Mg<jats:sup>2+</jats:sup>. Previous studies have shown that phosphorylation of a PhoP variant with a C-terminal His tag (PhoP<jats:sub>His</jats:sub>) enhances dimerization and binding to target DNA. Here, the effect of phosphorylation on the oligomerization and DNA binding properties of both wild-type PhoP (PhoP) and PhoP<jats:sub>His</jats:sub>are compared. Gel filtration chromatography showed that PhoP exists as a mixture of monomer and dimer regardless of its phosphorylation state. In contrast, unphosphorylated PhoP<jats:sub>His</jats:sub>was mostly monomeric, whereas PhoP<jats:sub>His</jats:sub>∼P existed as a mixture of monomer and dimer. By monitoring the tryptophan fluorescence of the proteins and the fluorescence of the probe 1-anilinonaphthalene-8-sulfonic acid bound to them, it was found that PhoP and PhoP<jats:sub>His</jats:sub>exhibited different spectral properties. The interaction between PhoP or PhoP<jats:sub>His</jats:sub>and the PhoP box of the<jats:italic>mgtA</jats:italic>promoter was monitored by surface plasmon resonance. Binding of PhoP to the PhoP box was barely influenced by phosphorylation. In contrast, phosphorylation of PhoP<jats:sub>His</jats:sub>clearly increased the interaction of PhoP<jats:sub>His</jats:sub>with target DNA. Altogether, these data show that a His tag at the C-terminus of PhoP affects its biochemical properties, most likely by affecting its conformation and/or its oligomerization state. More importantly, these results show that wild-type PhoP dimerization and interaction with target DNA are independent of phosphorylation, which is in contrast to the previously proposed model.</jats:p> |
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| imprint_str_mv | Microbiology Society, 2005 |
| institution | DE-Gla1, DE-Zi4, DE-15, DE-Rs1, DE-Pl11, DE-105, DE-14, DE-Ch1, DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161 |
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| series | Microbiology |
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| spelling | Perron-Savard, Philippe De Crescenzo, Gregory Moual, Hervé Le 1350-0872 1465-2080 Microbiology Society Microbiology http://dx.doi.org/10.1099/mic.0.28236-0 <jats:p>In<jats:italic>Salmonella enterica</jats:italic>, PhoP is the response regulator of the PhoP/PhoQ two-component regulatory system that controls the expression of various virulence factors in response to external Mg<jats:sup>2+</jats:sup>. Previous studies have shown that phosphorylation of a PhoP variant with a C-terminal His tag (PhoP<jats:sub>His</jats:sub>) enhances dimerization and binding to target DNA. Here, the effect of phosphorylation on the oligomerization and DNA binding properties of both wild-type PhoP (PhoP) and PhoP<jats:sub>His</jats:sub>are compared. Gel filtration chromatography showed that PhoP exists as a mixture of monomer and dimer regardless of its phosphorylation state. In contrast, unphosphorylated PhoP<jats:sub>His</jats:sub>was mostly monomeric, whereas PhoP<jats:sub>His</jats:sub>∼P existed as a mixture of monomer and dimer. By monitoring the tryptophan fluorescence of the proteins and the fluorescence of the probe 1-anilinonaphthalene-8-sulfonic acid bound to them, it was found that PhoP and PhoP<jats:sub>His</jats:sub>exhibited different spectral properties. The interaction between PhoP or PhoP<jats:sub>His</jats:sub>and the PhoP box of the<jats:italic>mgtA</jats:italic>promoter was monitored by surface plasmon resonance. Binding of PhoP to the PhoP box was barely influenced by phosphorylation. In contrast, phosphorylation of PhoP<jats:sub>His</jats:sub>clearly increased the interaction of PhoP<jats:sub>His</jats:sub>with target DNA. Altogether, these data show that a His tag at the C-terminus of PhoP affects its biochemical properties, most likely by affecting its conformation and/or its oligomerization state. More importantly, these results show that wild-type PhoP dimerization and interaction with target DNA are independent of phosphorylation, which is in contrast to the previously proposed model.</jats:p> Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent Microbiology |
| spellingShingle | Perron-Savard, Philippe, De Crescenzo, Gregory, Moual, Hervé Le, Microbiology, Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent, Microbiology |
| title | Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_full | Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_fullStr | Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_full_unstemmed | Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_short | Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| title_sort | dimerization and dna binding of the salmonella enterica phop response regulator are phosphorylation independent |
| title_unstemmed | Dimerization and DNA binding of the Salmonella enterica PhoP response regulator are phosphorylation independent |
| topic | Microbiology |
| url | http://dx.doi.org/10.1099/mic.0.28236-0 |