Transcript counting in single cells reveals dynamics of rDNA transcription
Most eukaryotes contain many tandem repeats of ribosomal RNA genes of which only a subset is transcribed at any given time. Current biochemical methods allow for the determination of the fraction of transcribing repeats (ON) versus non-transcribing repeats (OFF) but do not provide any dynamical info...
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2011
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| Online Access: | http://hdl.handle.net/1721.1/61692 |
| _version_ | 1826216728935268352 |
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| author | Tan, Rui Zhen van Oudenaarden, Alexander |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Tan, Rui Zhen van Oudenaarden, Alexander |
| author_sort | Tan, Rui Zhen |
| collection | MIT |
| description | Most eukaryotes contain many tandem repeats of ribosomal RNA genes of which only a subset is transcribed at any given time. Current biochemical methods allow for the determination of the fraction of transcribing repeats (ON) versus non-transcribing repeats (OFF) but do not provide any dynamical information and obscure any transcription activity at the single-cell level. Here, we use a fluorescence in situ hybridization (FISH) technique that allows the detection of single-RNA molecules in individual yeast cells. We use this method complemented with theoretical modeling to determine the rate of switching from OFF to ON (activation rate) and the average number of RNA molecules produced during each transcriptional burst (burst size). We explore how these two variables change in mutants and different growth conditions, and show that this method resolves changes in these two variables even when the average rDNA expression is unaltered. These phenotypic changes could not have been detected by traditional biochemical assays. |
| first_indexed | 2024-09-23T16:52:09Z |
| format | Article |
| id | mit-1721.1/61692 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:52:09Z |
| publishDate | 2011 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/616922022-10-03T08:49:00Z Transcript counting in single cells reveals dynamics of rDNA transcription Tan, Rui Zhen van Oudenaarden, Alexander Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Physics van Oudenaarden, Alexander van Oudenaarden, Alexander Most eukaryotes contain many tandem repeats of ribosomal RNA genes of which only a subset is transcribed at any given time. Current biochemical methods allow for the determination of the fraction of transcribing repeats (ON) versus non-transcribing repeats (OFF) but do not provide any dynamical information and obscure any transcription activity at the single-cell level. Here, we use a fluorescence in situ hybridization (FISH) technique that allows the detection of single-RNA molecules in individual yeast cells. We use this method complemented with theoretical modeling to determine the rate of switching from OFF to ON (activation rate) and the average number of RNA molecules produced during each transcriptional burst (burst size). We explore how these two variables change in mutants and different growth conditions, and show that this method resolves changes in these two variables even when the average rDNA expression is unaltered. These phenotypic changes could not have been detected by traditional biochemical assays. National Science Foundation (U.S.) (Grant No. NSF-0548484) National Institutes of Health (U.S) (R01-GM068957) 2011-03-14T19:09:54Z 2011-03-14T19:09:54Z 2010-04 2009-08 Article http://purl.org/eprint/type/JournalArticle 1744-4292 http://hdl.handle.net/1721.1/61692 Tan, Rui Zhen, and Alexander van Oudenaarden. “Transcript counting in single cells reveals dynamics of rDNA transcription.”Molecular Systems Biology 6(2010):358 © 2010 Nature Publishing Group en_US http://dx.doi.org/10.1038/msb.2010.14 Molecular Systems Biology Creative Commons Attribution-Non-Commercial-Share Alike 3.0 http://creativecommons.org/licenses/by/3.0 application/pdf Nature Publishing Group Molecular Systems Biology |
| spellingShingle | Tan, Rui Zhen van Oudenaarden, Alexander Transcript counting in single cells reveals dynamics of rDNA transcription |
| title | Transcript counting in single cells reveals dynamics of rDNA transcription |
| title_full | Transcript counting in single cells reveals dynamics of rDNA transcription |
| title_fullStr | Transcript counting in single cells reveals dynamics of rDNA transcription |
| title_full_unstemmed | Transcript counting in single cells reveals dynamics of rDNA transcription |
| title_short | Transcript counting in single cells reveals dynamics of rDNA transcription |
| title_sort | transcript counting in single cells reveals dynamics of rdna transcription |
| url | http://hdl.handle.net/1721.1/61692 |
| work_keys_str_mv | AT tanruizhen transcriptcountinginsinglecellsrevealsdynamicsofrdnatranscription AT vanoudenaardenalexander transcriptcountinginsinglecellsrevealsdynamicsofrdnatranscription |