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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
32
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pubmed:dateCreated |
2009-8-26
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pubmed:abstractText |
A long-standing question in retrovirus biology is how RNA genomes are distributed among virions. In the studies presented in this report, we addressed this issue by directly examining HIV-1 RNAs in virions using a modified HIV-1 genome that contained recognition sites for BglG, an antitermination protein in the Escherichia coli bgl operon, which was coexpressed with a fragment of BglG RNA binding protein fused to a fluorescent protein. Our results demonstrate that the majority of virions (>90%) contain viral RNAs. We also coexpressed HIV-1 genomes containing binding sites for BglG or the bacteriophage MS2 coat protein along with 2 fluorescent protein-tagged RNA binding proteins. This method allows simultaneously labeling and discrimination of 2 different RNAs at single-RNA-detection sensitivity. Using this strategy, we obtained physical evidence that virions contain RNAs derived from different parental viruses (heterozygous virion) at ratios expected from a random distribution, and we found that this ratio can be altered by changing the dimerization sequences. Our studies of heterozygous virions also support a generally accepted but unproven assumption that most particles contain 1 dimer. This study provides answers to long-standing questions in HIV-1 biology and illustrates the power and sensitivity of the 2-RNA labeling method, which can also be adapted to analyze various issues of RNA biogenesis including the detection of different RNAs in live cell imaging.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-11320254,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-12546792,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-12663764,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-12852860,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-14512567,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-14668432,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-15650192,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-15956186,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-16230638,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-1698125,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-17267488,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-17267494,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-17392359,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-17599354,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-17954611,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-18500329,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-19064259,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-7824947,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-8096089,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-8510214,
http://linkedlifedata.com/resource/pubmed/commentcorrection/19628694-9809065
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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 |
Aug
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pubmed:issn |
1091-6490
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:day |
11
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pubmed:volume |
106
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
13535-40
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pubmed:dateRevised |
2010-8-12
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pubmed:meshHeading |
pubmed-meshheading:19628694-Bacterial Proteins,
pubmed-meshheading:19628694-Base Pairing,
pubmed-meshheading:19628694-Base Sequence,
pubmed-meshheading:19628694-Capsid Proteins,
pubmed-meshheading:19628694-Cell Line,
pubmed-meshheading:19628694-Fluorescence,
pubmed-meshheading:19628694-Genome, Viral,
pubmed-meshheading:19628694-HIV-1,
pubmed-meshheading:19628694-Heterozygote,
pubmed-meshheading:19628694-Homozygote,
pubmed-meshheading:19628694-Humans,
pubmed-meshheading:19628694-Levivirus,
pubmed-meshheading:19628694-Molecular Sequence Data,
pubmed-meshheading:19628694-Nucleic Acid Conformation,
pubmed-meshheading:19628694-Operon,
pubmed-meshheading:19628694-RNA, Viral,
pubmed-meshheading:19628694-RNA-Binding Proteins,
pubmed-meshheading:19628694-Staining and Labeling,
pubmed-meshheading:19628694-Virion,
pubmed-meshheading:19628694-Virus Assembly
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pubmed:year |
2009
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pubmed:articleTitle |
High efficiency of HIV-1 genomic RNA packaging and heterozygote formation revealed by single virion analysis.
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pubmed:affiliation |
HIV Drug Resistance Program, National Cancer Institute, Frederick, MD 21702, USA.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural,
Research Support, N.I.H., Intramural
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