Characterization of sequence elements from Malvastrum yellow vein betasatellite regulating promoter activity and DNA replication.
Identifieur interne : 000488 ( Main/Corpus ); précédent : 000487; suivant : 000489Characterization of sequence elements from Malvastrum yellow vein betasatellite regulating promoter activity and DNA replication.
Auteurs : Jie Zhang ; Xinyue Zhang ; Yaqin Wang ; Huwei Hou ; Yajuan QianSource :
- Virology journal [ 1743-422X ] ; 2012.
English descriptors
- KwdEn :
- Base Sequence (MeSH), DNA Mutational Analysis (MeSH), DNA Replication (MeSH), DNA, Satellite (genetics), Genes, Reporter (MeSH), Glucuronidase (analysis), Glucuronidase (genetics), Malvaceae (virology), Molecular Sequence Data (MeSH), Plant Diseases (virology), Promoter Regions, Genetic (MeSH), Tobacco (virology).
- MESH :
- chemical , analysis : Glucuronidase.
- chemical , genetics : DNA, Satellite, Glucuronidase.
- virology : Malvaceae, Plant Diseases, Tobacco.
- Base Sequence, DNA Mutational Analysis, DNA Replication, Genes, Reporter, Molecular Sequence Data, Promoter Regions, Genetic.
Abstract
BACKGROUND
Many monopartite begomoviruses are associated with betasatellites, but only several promoters from which were isolated and studied. In this study, the βC1 promoter from Malvastrum yellow vein betasatellite (MYVB) was characterized and important sequence elements were identified to modulate promoter activity and replication of MYVB.
RESULTS
A 991 nucleotide (nt) fragment upstream of the translation start site of the βC1 open reading frame of MYVB and a series of deletions within this fragment were constructed and fused to the β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes, respectively. Agrobacterium-mediated transient expression assays showed that the 991 nt fragment was functional and that a 28 nt region (between -390 nt and -418 nt), which includes a 5'UTR Py-rich stretch motif, was important for promoter activity. Replication assays using Nicotiana benthamiana leaf discs and whole plants showed that deletion of the 5'UTR Py-rich stretch impaired viral satellite replication in the presence of the helper virus. Transgenic assays demonstrated that the 991 nt fragment conferred a constitutive expression pattern in transgenic tobacco plants and that a 214 nt fragment at the 3'-end of this sequence was sufficient to drive this expression pattern.
CONCLUSION
Our results showed that the βC1 promoter of MYVB displayed a constitutive expression pattern and a 5'UTR Py-rich stretch motif regulated both βC1 promoter activity and MYVB replication.
DOI: 10.1186/1743-422X-9-234
PubMed: 23057573
PubMed Central: PMC3544650
Links to Exploration step
pubmed:23057573Le document en format XML
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<author><name sortKey="Zhang, Jie" sort="Zhang, Jie" uniqKey="Zhang J" first="Jie" last="Zhang">Jie Zhang</name>
<affiliation><nlm:affiliation>State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, People's Republic of China.</nlm:affiliation>
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<author><name sortKey="Zhang, Xinyue" sort="Zhang, Xinyue" uniqKey="Zhang X" first="Xinyue" last="Zhang">Xinyue Zhang</name>
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<author><name sortKey="Wang, Yaqin" sort="Wang, Yaqin" uniqKey="Wang Y" first="Yaqin" last="Wang">Yaqin Wang</name>
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<author><name sortKey="Hou, Huwei" sort="Hou, Huwei" uniqKey="Hou H" first="Huwei" last="Hou">Huwei Hou</name>
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<author><name sortKey="Qian, Yajuan" sort="Qian, Yajuan" uniqKey="Qian Y" first="Yajuan" last="Qian">Yajuan Qian</name>
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<author><name sortKey="Zhang, Jie" sort="Zhang, Jie" uniqKey="Zhang J" first="Jie" last="Zhang">Jie Zhang</name>
<affiliation><nlm:affiliation>State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, People's Republic of China.</nlm:affiliation>
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<author><name sortKey="Zhang, Xinyue" sort="Zhang, Xinyue" uniqKey="Zhang X" first="Xinyue" last="Zhang">Xinyue Zhang</name>
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<author><name sortKey="Wang, Yaqin" sort="Wang, Yaqin" uniqKey="Wang Y" first="Yaqin" last="Wang">Yaqin Wang</name>
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<author><name sortKey="Hou, Huwei" sort="Hou, Huwei" uniqKey="Hou H" first="Huwei" last="Hou">Huwei Hou</name>
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<author><name sortKey="Qian, Yajuan" sort="Qian, Yajuan" uniqKey="Qian Y" first="Yajuan" last="Qian">Yajuan Qian</name>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence (MeSH)</term>
<term>DNA Mutational Analysis (MeSH)</term>
<term>DNA Replication (MeSH)</term>
<term>DNA, Satellite (genetics)</term>
<term>Genes, Reporter (MeSH)</term>
<term>Glucuronidase (analysis)</term>
<term>Glucuronidase (genetics)</term>
<term>Malvaceae (virology)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Plant Diseases (virology)</term>
<term>Promoter Regions, Genetic (MeSH)</term>
<term>Tobacco (virology)</term>
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<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Glucuronidase</term>
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<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Satellite</term>
<term>Glucuronidase</term>
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<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Malvaceae</term>
<term>Plant Diseases</term>
<term>Tobacco</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term>
<term>DNA Mutational Analysis</term>
<term>DNA Replication</term>
<term>Genes, Reporter</term>
<term>Molecular Sequence Data</term>
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<front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b>
</p>
<p>Many monopartite begomoviruses are associated with betasatellites, but only several promoters from which were isolated and studied. In this study, the βC1 promoter from Malvastrum yellow vein betasatellite (MYVB) was characterized and important sequence elements were identified to modulate promoter activity and replication of MYVB.</p>
</div>
<div type="abstract" xml:lang="en"><p><b>RESULTS</b>
</p>
<p>A 991 nucleotide (nt) fragment upstream of the translation start site of the βC1 open reading frame of MYVB and a series of deletions within this fragment were constructed and fused to the β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes, respectively. Agrobacterium-mediated transient expression assays showed that the 991 nt fragment was functional and that a 28 nt region (between -390 nt and -418 nt), which includes a 5'UTR Py-rich stretch motif, was important for promoter activity. Replication assays using Nicotiana benthamiana leaf discs and whole plants showed that deletion of the 5'UTR Py-rich stretch impaired viral satellite replication in the presence of the helper virus. Transgenic assays demonstrated that the 991 nt fragment conferred a constitutive expression pattern in transgenic tobacco plants and that a 214 nt fragment at the 3'-end of this sequence was sufficient to drive this expression pattern.</p>
</div>
<div type="abstract" xml:lang="en"><p><b>CONCLUSION</b>
</p>
<p>Our results showed that the βC1 promoter of MYVB displayed a constitutive expression pattern and a 5'UTR Py-rich stretch motif regulated both βC1 promoter activity and MYVB replication.</p>
</div>
</front>
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<Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">Many monopartite begomoviruses are associated with betasatellites, but only several promoters from which were isolated and studied. In this study, the βC1 promoter from Malvastrum yellow vein betasatellite (MYVB) was characterized and important sequence elements were identified to modulate promoter activity and replication of MYVB.</AbstractText>
<AbstractText Label="RESULTS" NlmCategory="RESULTS">A 991 nucleotide (nt) fragment upstream of the translation start site of the βC1 open reading frame of MYVB and a series of deletions within this fragment were constructed and fused to the β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes, respectively. Agrobacterium-mediated transient expression assays showed that the 991 nt fragment was functional and that a 28 nt region (between -390 nt and -418 nt), which includes a 5'UTR Py-rich stretch motif, was important for promoter activity. Replication assays using Nicotiana benthamiana leaf discs and whole plants showed that deletion of the 5'UTR Py-rich stretch impaired viral satellite replication in the presence of the helper virus. Transgenic assays demonstrated that the 991 nt fragment conferred a constitutive expression pattern in transgenic tobacco plants and that a 214 nt fragment at the 3'-end of this sequence was sufficient to drive this expression pattern.</AbstractText>
<AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">Our results showed that the βC1 promoter of MYVB displayed a constitutive expression pattern and a 5'UTR Py-rich stretch motif regulated both βC1 promoter activity and MYVB replication.</AbstractText>
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