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Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR
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| Zeitschriftentitel: | Journal of Bacteriology |
|---|---|
| Personen und Körperschaften: | , , , , |
| In: | Journal of Bacteriology, 189, 2007, 23, S. 8651-8659 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Society for Microbiology
|
| Schlagwörter: |
| author_facet |
Wang, Xueqi Cheng, Zhihui Zhang, Chunbin Kikuchi, Takane Rikihisa, Yasuko Wang, Xueqi Cheng, Zhihui Zhang, Chunbin Kikuchi, Takane Rikihisa, Yasuko |
|---|---|
| author |
Wang, Xueqi Cheng, Zhihui Zhang, Chunbin Kikuchi, Takane Rikihisa, Yasuko |
| spellingShingle |
Wang, Xueqi Cheng, Zhihui Zhang, Chunbin Kikuchi, Takane Rikihisa, Yasuko Journal of Bacteriology Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR Molecular Biology Microbiology |
| author_sort |
wang, xueqi |
| spelling |
Wang, Xueqi Cheng, Zhihui Zhang, Chunbin Kikuchi, Takane Rikihisa, Yasuko 0021-9193 1098-5530 American Society for Microbiology Molecular Biology Microbiology http://dx.doi.org/10.1128/jb.00881-07 <jats:title>ABSTRACT</jats:title> <jats:p> The natural life cycle of <jats:italic>Anaplasma phagocytophilum</jats:italic> , an obligatory intracellular bacterium that causes human granulocytic anaplasmosis, consists of alternate infection of two distinct hosts, ticks and mammals, in which bacterial surface proteins are expected to have a critical role. The present study investigated regulation of <jats:italic>A. phagocytophilum p44</jats:italic> genes, which encode the P44 major surface proteins. Quantitative real-time reverse transcription-PCR analysis revealed that the amount of <jats:italic>p44</jats:italic> mRNA obtained from spleens of <jats:italic>A. phagocytophilum</jats:italic> -infected SCID mice was approximately 10-fold greater than the amount obtained from salivary glands of <jats:italic>A. phagocytophilum</jats:italic> -infected <jats:italic>Ixodes scapularis</jats:italic> nymphs. Similarly, the amount of <jats:italic>p44</jats:italic> mRNA obtained from <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells per bacterium was significantly greater than the amount obtained from infected ISE6 tick cells. The relative amount of <jats:italic>p44</jats:italic> mRNA was approximately threefold higher in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells cultured at 37°C than in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells cultured at 28°C. Although there are more than 100 <jats:italic>p44</jats:italic> paralogs, we observed expression mainly from the <jats:italic>p44</jats:italic> expression locus ( <jats:italic>p44E</jats:italic> ) in various host environments. Interestingly, transcription of the <jats:italic>A. phagocytophilum</jats:italic> gene encoding the DNA binding protein ApxR was also significantly greater in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells than in infected ISE6 tick cells. Gel mobility shift and DNase I protection assays revealed recombinant ApxR binding to the promoter regions of <jats:italic>p44E</jats:italic> and <jats:italic>apxR</jats:italic> . ApxR also transactivated the <jats:italic>p44E</jats:italic> and <jats:italic>apxR</jats:italic> promoter regions in a <jats:italic>lacZ</jats:italic> reporter assay. These results indicate that <jats:italic>p44</jats:italic> genes and <jats:italic>apxR</jats:italic> are specifically up-regulated in the mammalian host environment and suggest that ApxR not only is positively autoregulated but also acts as a transcriptional regulator of <jats:italic>p44E</jats:italic> . </jats:p> <i>Anaplasma phagocytophilum p44</i> mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR Journal of Bacteriology |
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10.1128/jb.00881-07 |
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American Society for Microbiology, 2007 |
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American Society for Microbiology, 2007 |
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0021-9193 1098-5530 |
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2007 |
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American Society for Microbiology |
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Journal of Bacteriology |
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49 |
| title |
Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_unstemmed |
Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_full |
Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_fullStr |
Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_full_unstemmed |
Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_short |
Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_sort |
<i>anaplasma phagocytophilum p44</i>
mrna expression is differentially regulated in mammalian and tick host cells: involvement of the dna binding protein apxr |
| topic |
Molecular Biology Microbiology |
| url |
http://dx.doi.org/10.1128/jb.00881-07 |
| publishDate |
2007 |
| physical |
8651-8659 |
| description |
<jats:title>ABSTRACT</jats:title>
<jats:p>
The natural life cycle of
<jats:italic>Anaplasma phagocytophilum</jats:italic>
, an obligatory intracellular bacterium that causes human granulocytic anaplasmosis, consists of alternate infection of two distinct hosts, ticks and mammals, in which bacterial surface proteins are expected to have a critical role. The present study investigated regulation of
<jats:italic>A. phagocytophilum p44</jats:italic>
genes, which encode the P44 major surface proteins. Quantitative real-time reverse transcription-PCR analysis revealed that the amount of
<jats:italic>p44</jats:italic>
mRNA obtained from spleens of
<jats:italic>A. phagocytophilum</jats:italic>
-infected SCID mice was approximately 10-fold greater than the amount obtained from salivary glands of
<jats:italic>A. phagocytophilum</jats:italic>
-infected
<jats:italic>Ixodes scapularis</jats:italic>
nymphs. Similarly, the amount of
<jats:italic>p44</jats:italic>
mRNA obtained from
<jats:italic>A. phagocytophilum</jats:italic>
-infected HL-60 cells per bacterium was significantly greater than the amount obtained from infected ISE6 tick cells. The relative amount of
<jats:italic>p44</jats:italic>
mRNA was approximately threefold higher in
<jats:italic>A. phagocytophilum</jats:italic>
-infected HL-60 cells cultured at 37°C than in
<jats:italic>A. phagocytophilum</jats:italic>
-infected HL-60 cells cultured at 28°C. Although there are more than 100
<jats:italic>p44</jats:italic>
paralogs, we observed expression mainly from the
<jats:italic>p44</jats:italic>
expression locus (
<jats:italic>p44E</jats:italic>
) in various host environments. Interestingly, transcription of the
<jats:italic>A. phagocytophilum</jats:italic>
gene encoding the DNA binding protein ApxR was also significantly greater in
<jats:italic>A. phagocytophilum</jats:italic>
-infected HL-60 cells than in infected ISE6 tick cells. Gel mobility shift and DNase I protection assays revealed recombinant ApxR binding to the promoter regions of
<jats:italic>p44E</jats:italic>
and
<jats:italic>apxR</jats:italic>
. ApxR also transactivated the
<jats:italic>p44E</jats:italic>
and
<jats:italic>apxR</jats:italic>
promoter regions in a
<jats:italic>lacZ</jats:italic>
reporter assay. These results indicate that
<jats:italic>p44</jats:italic>
genes and
<jats:italic>apxR</jats:italic>
are specifically up-regulated in the mammalian host environment and suggest that ApxR not only is positively autoregulated but also acts as a transcriptional regulator of
<jats:italic>p44E</jats:italic>
.
</jats:p> |
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| description | <jats:title>ABSTRACT</jats:title> <jats:p> The natural life cycle of <jats:italic>Anaplasma phagocytophilum</jats:italic> , an obligatory intracellular bacterium that causes human granulocytic anaplasmosis, consists of alternate infection of two distinct hosts, ticks and mammals, in which bacterial surface proteins are expected to have a critical role. The present study investigated regulation of <jats:italic>A. phagocytophilum p44</jats:italic> genes, which encode the P44 major surface proteins. Quantitative real-time reverse transcription-PCR analysis revealed that the amount of <jats:italic>p44</jats:italic> mRNA obtained from spleens of <jats:italic>A. phagocytophilum</jats:italic> -infected SCID mice was approximately 10-fold greater than the amount obtained from salivary glands of <jats:italic>A. phagocytophilum</jats:italic> -infected <jats:italic>Ixodes scapularis</jats:italic> nymphs. Similarly, the amount of <jats:italic>p44</jats:italic> mRNA obtained from <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells per bacterium was significantly greater than the amount obtained from infected ISE6 tick cells. The relative amount of <jats:italic>p44</jats:italic> mRNA was approximately threefold higher in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells cultured at 37°C than in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells cultured at 28°C. Although there are more than 100 <jats:italic>p44</jats:italic> paralogs, we observed expression mainly from the <jats:italic>p44</jats:italic> expression locus ( <jats:italic>p44E</jats:italic> ) in various host environments. Interestingly, transcription of the <jats:italic>A. phagocytophilum</jats:italic> gene encoding the DNA binding protein ApxR was also significantly greater in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells than in infected ISE6 tick cells. Gel mobility shift and DNase I protection assays revealed recombinant ApxR binding to the promoter regions of <jats:italic>p44E</jats:italic> and <jats:italic>apxR</jats:italic> . ApxR also transactivated the <jats:italic>p44E</jats:italic> and <jats:italic>apxR</jats:italic> promoter regions in a <jats:italic>lacZ</jats:italic> reporter assay. These results indicate that <jats:italic>p44</jats:italic> genes and <jats:italic>apxR</jats:italic> are specifically up-regulated in the mammalian host environment and suggest that ApxR not only is positively autoregulated but also acts as a transcriptional regulator of <jats:italic>p44E</jats:italic> . </jats:p> |
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| spelling | Wang, Xueqi Cheng, Zhihui Zhang, Chunbin Kikuchi, Takane Rikihisa, Yasuko 0021-9193 1098-5530 American Society for Microbiology Molecular Biology Microbiology http://dx.doi.org/10.1128/jb.00881-07 <jats:title>ABSTRACT</jats:title> <jats:p> The natural life cycle of <jats:italic>Anaplasma phagocytophilum</jats:italic> , an obligatory intracellular bacterium that causes human granulocytic anaplasmosis, consists of alternate infection of two distinct hosts, ticks and mammals, in which bacterial surface proteins are expected to have a critical role. The present study investigated regulation of <jats:italic>A. phagocytophilum p44</jats:italic> genes, which encode the P44 major surface proteins. Quantitative real-time reverse transcription-PCR analysis revealed that the amount of <jats:italic>p44</jats:italic> mRNA obtained from spleens of <jats:italic>A. phagocytophilum</jats:italic> -infected SCID mice was approximately 10-fold greater than the amount obtained from salivary glands of <jats:italic>A. phagocytophilum</jats:italic> -infected <jats:italic>Ixodes scapularis</jats:italic> nymphs. Similarly, the amount of <jats:italic>p44</jats:italic> mRNA obtained from <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells per bacterium was significantly greater than the amount obtained from infected ISE6 tick cells. The relative amount of <jats:italic>p44</jats:italic> mRNA was approximately threefold higher in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells cultured at 37°C than in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells cultured at 28°C. Although there are more than 100 <jats:italic>p44</jats:italic> paralogs, we observed expression mainly from the <jats:italic>p44</jats:italic> expression locus ( <jats:italic>p44E</jats:italic> ) in various host environments. Interestingly, transcription of the <jats:italic>A. phagocytophilum</jats:italic> gene encoding the DNA binding protein ApxR was also significantly greater in <jats:italic>A. phagocytophilum</jats:italic> -infected HL-60 cells than in infected ISE6 tick cells. Gel mobility shift and DNase I protection assays revealed recombinant ApxR binding to the promoter regions of <jats:italic>p44E</jats:italic> and <jats:italic>apxR</jats:italic> . ApxR also transactivated the <jats:italic>p44E</jats:italic> and <jats:italic>apxR</jats:italic> promoter regions in a <jats:italic>lacZ</jats:italic> reporter assay. These results indicate that <jats:italic>p44</jats:italic> genes and <jats:italic>apxR</jats:italic> are specifically up-regulated in the mammalian host environment and suggest that ApxR not only is positively autoregulated but also acts as a transcriptional regulator of <jats:italic>p44E</jats:italic> . </jats:p> <i>Anaplasma phagocytophilum p44</i> mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR Journal of Bacteriology |
| spellingShingle | Wang, Xueqi, Cheng, Zhihui, Zhang, Chunbin, Kikuchi, Takane, Rikihisa, Yasuko, Journal of Bacteriology, Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR, Molecular Biology, Microbiology |
| title | Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_full | Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_fullStr | Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_full_unstemmed | Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_short | Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| title_sort | <i>anaplasma phagocytophilum p44</i> mrna expression is differentially regulated in mammalian and tick host cells: involvement of the dna binding protein apxr |
| title_unstemmed | Anaplasma phagocytophilum p44 mRNA Expression Is Differentially Regulated in Mammalian and Tick Host Cells: Involvement of the DNA Binding Protein ApxR |
| topic | Molecular Biology, Microbiology |
| url | http://dx.doi.org/10.1128/jb.00881-07 |