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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
8
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pubmed:dateCreated |
2001-3-26
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pubmed:abstractText |
The Sindbis virus minimal subgenomic mRNA promoter (spanning positions -19 to +5 relative to the subgenomic mRNA start site) is approximately three- to sixfold less active than the fully active -98 to +14 promoter region. We identified two elements flanking the -19 to +5 region which increase its transcription to levels comparable to the -98 to +14 region. These elements span positions -40 to -20 and +6 to +14 and act synergistically to enhance transcription. Nine different virus libraries were constructed containing blocks of five randomized nucleotides at various positions in the -40 to +14 region. On passaging these libraries in mosquito cells, a small subset of the viruses came to dominate the population. Sequence analysis at the population level and for individual clones revealed that in general, wild-type bases were preferred for positions -15 to +5 of the minimal promoter. Base mutagenesis experiments indicated that the selection of wild-type bases in this region was primarily due to requirements for subgenomic mRNA transcription. Outside of the minimal promoter, the -35 to -29 region contained four positions which also preferred wildtype bases. However, the remaining positions generally preferred non-wild-type bases. On passaging of the virus libraries on hamster cells, the -15 to +5 region again preferred the wild-type base but most of the remaining positions exhibited almost no base preference. The promoter thus consists of an essential central region from -15 to +5 and discrete flanking sites that render it fully active, depending on the host environment.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-10191183,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-10590104,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-10640553,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-10769078,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-11070017,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-1309918,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-1370252,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-196393,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2016769,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2053283,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2072446,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2155505,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2319643,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2319651,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2384916,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2440030,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2685355,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2834873,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-2886548,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-3088830,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-3318675,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-3334750,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-3488539,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-3753584,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-3973983,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-6154866,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-6259811,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-6288962,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-6291034,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-6308269,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-7494287,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-7494288,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-7817880,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-7968923,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-8317146,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-8806532,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-8892905,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-9007072,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-9032308,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-9188557,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-9343230,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-9371567,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11264340-9875334
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0022-538X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
75
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
3509-19
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:11264340-Amino Acid Sequence,
pubmed-meshheading:11264340-Animals,
pubmed-meshheading:11264340-Base Sequence,
pubmed-meshheading:11264340-Cell Line,
pubmed-meshheading:11264340-Cloning, Molecular,
pubmed-meshheading:11264340-Consensus Sequence,
pubmed-meshheading:11264340-Cricetinae,
pubmed-meshheading:11264340-Gene Library,
pubmed-meshheading:11264340-Genome, Viral,
pubmed-meshheading:11264340-Molecular Sequence Data,
pubmed-meshheading:11264340-Point Mutation,
pubmed-meshheading:11264340-Promoter Regions, Genetic,
pubmed-meshheading:11264340-RNA, Messenger,
pubmed-meshheading:11264340-RNA, Viral,
pubmed-meshheading:11264340-Sequence Alignment,
pubmed-meshheading:11264340-Sequence Deletion,
pubmed-meshheading:11264340-Serial Passage,
pubmed-meshheading:11264340-Sindbis Virus,
pubmed-meshheading:11264340-Transcription, Genetic,
pubmed-meshheading:11264340-Transfection
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pubmed:year |
2001
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pubmed:articleTitle |
Sequence requirements for Sindbis virus subgenomic mRNA promoter function in cultured cells.
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pubmed:affiliation |
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110-1093, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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