A Novel Regulator PepR Regulates the Expression of Dipeptidase Gene <i>pepV</i> in <i>Bacillus thuringiensis</i>
<i>Bacillus thuringiensis</i> produces insecticidal crystal proteins encoded by <i>cry</i> or <i>cyt</i> genes and targets a variety of insect pests. We previously found that a strong promoter of a DeoR family transcriptional regulator (HD73_5014) can efficiently...
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MDPI AG
2024-03-01
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| Series: | Microorganisms |
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| author | Xin Zhang Hengjie Wang Tinglu Yan Yuhan Chen Qi Peng Fuping Song |
| author_facet | Xin Zhang Hengjie Wang Tinglu Yan Yuhan Chen Qi Peng Fuping Song |
| author_sort | Xin Zhang |
| collection | DOAJ |
| description | <i>Bacillus thuringiensis</i> produces insecticidal crystal proteins encoded by <i>cry</i> or <i>cyt</i> genes and targets a variety of insect pests. We previously found that a strong promoter of a DeoR family transcriptional regulator (HD73_5014) can efficiently drive <i>cry1Ac</i> expression in <i>B. thuringiensis</i> HD73. Here, we investigated the regulation of neighbor genes by HD73_5014. The HD73_5014 homologs are widely distributed in Gram-positive bacterial species. Its neighbor genes include <i>pepV</i>, <i>rsuA</i>, and <i>ytgP</i>, which encode dipeptidase, rRNA pseudouridine synthase and polysaccharide biosynthesis protein, respectively. The four open reading frames (ORFs) are organized to be a <i>pepR</i> gene cluster in HD73. RT-PCR analysis revealed that the <i>rsuA</i> and <i>ytgP</i> genes formed a transcriptional unit (<i>rsuA</i>-<i>ytgP</i> operon), while <i>pepV</i> formed a transcriptional unit in HD73. Promoter-<i>lacZ</i> fusion assays showed that the <i>pepV</i> and <i>rsuA</i>-<i>ytgP</i> promoters are regulated by HD73_5014. EMSA experiments showed that HD73_5014 directly binds to the <i>pepV</i> promoter region but not to the <i>rusA</i>-<i>ytgP</i> promoter region. Thus, the HD73_5014 transcriptional regulator, which controls the expression of the dipeptidase <i>pepV</i>, was named PepR (dipeptidase regulator). We also confirmed the direct regulation between PepR and PepV by the increased sensitivity to vancomycin in Δ<i>pepV</i> and Δ<i>pepR</i> mutants compared to HD73. |
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| language | English |
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| publishDate | 2024-03-01 |
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| spelling | doaj.art-30f024ff07ea4d5584a30e1a69d6a90a2024-03-27T13:55:53ZengMDPI AGMicroorganisms2076-26072024-03-0112357910.3390/microorganisms12030579A Novel Regulator PepR Regulates the Expression of Dipeptidase Gene <i>pepV</i> in <i>Bacillus thuringiensis</i>Xin Zhang0Hengjie Wang1Tinglu Yan2Yuhan Chen3Qi Peng4Fuping Song5State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China<i>Bacillus thuringiensis</i> produces insecticidal crystal proteins encoded by <i>cry</i> or <i>cyt</i> genes and targets a variety of insect pests. We previously found that a strong promoter of a DeoR family transcriptional regulator (HD73_5014) can efficiently drive <i>cry1Ac</i> expression in <i>B. thuringiensis</i> HD73. Here, we investigated the regulation of neighbor genes by HD73_5014. The HD73_5014 homologs are widely distributed in Gram-positive bacterial species. Its neighbor genes include <i>pepV</i>, <i>rsuA</i>, and <i>ytgP</i>, which encode dipeptidase, rRNA pseudouridine synthase and polysaccharide biosynthesis protein, respectively. The four open reading frames (ORFs) are organized to be a <i>pepR</i> gene cluster in HD73. RT-PCR analysis revealed that the <i>rsuA</i> and <i>ytgP</i> genes formed a transcriptional unit (<i>rsuA</i>-<i>ytgP</i> operon), while <i>pepV</i> formed a transcriptional unit in HD73. Promoter-<i>lacZ</i> fusion assays showed that the <i>pepV</i> and <i>rsuA</i>-<i>ytgP</i> promoters are regulated by HD73_5014. EMSA experiments showed that HD73_5014 directly binds to the <i>pepV</i> promoter region but not to the <i>rusA</i>-<i>ytgP</i> promoter region. Thus, the HD73_5014 transcriptional regulator, which controls the expression of the dipeptidase <i>pepV</i>, was named PepR (dipeptidase regulator). We also confirmed the direct regulation between PepR and PepV by the increased sensitivity to vancomycin in Δ<i>pepV</i> and Δ<i>pepR</i> mutants compared to HD73.https://www.mdpi.com/2076-2607/12/3/579transcriptional regulator PepRdipeptidase gene <i>pepV</i>resistant vancomycin resistance<i>Bacillus thuringiensis</i> |
| spellingShingle | Xin Zhang Hengjie Wang Tinglu Yan Yuhan Chen Qi Peng Fuping Song A Novel Regulator PepR Regulates the Expression of Dipeptidase Gene <i>pepV</i> in <i>Bacillus thuringiensis</i> Microorganisms transcriptional regulator PepR dipeptidase gene <i>pepV</i> resistant vancomycin resistance <i>Bacillus thuringiensis</i> |
| title | A Novel Regulator PepR Regulates the Expression of Dipeptidase Gene <i>pepV</i> in <i>Bacillus thuringiensis</i> |
| title_full | A Novel Regulator PepR Regulates the Expression of Dipeptidase Gene <i>pepV</i> in <i>Bacillus thuringiensis</i> |
| title_fullStr | A Novel Regulator PepR Regulates the Expression of Dipeptidase Gene <i>pepV</i> in <i>Bacillus thuringiensis</i> |
| title_full_unstemmed | A Novel Regulator PepR Regulates the Expression of Dipeptidase Gene <i>pepV</i> in <i>Bacillus thuringiensis</i> |
| title_short | A Novel Regulator PepR Regulates the Expression of Dipeptidase Gene <i>pepV</i> in <i>Bacillus thuringiensis</i> |
| title_sort | novel regulator pepr regulates the expression of dipeptidase gene i pepv i in i bacillus thuringiensis i |
| topic | transcriptional regulator PepR dipeptidase gene <i>pepV</i> resistant vancomycin resistance <i>Bacillus thuringiensis</i> |
| url | https://www.mdpi.com/2076-2607/12/3/579 |
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