Real-Time Observation of Transcription Initiation and Elongation on an Endogenous Yeast Gene
Cellular messenger RNA levels are achieved by the combinatorial complexity of factors controlling transcription, yet the small number of molecules involved in these pathways fluctuates stochastically. It has not yet been experimentally possible to observe the activity of single polymerases on an end...
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| Veröffentlicht in: | Science (American Association for the Advancement of Science) 2011-04, Vol.332 (6028), p.475-478 |
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| creator | Larson, Daniel R. Zenklusen, Daniel Wu, Bin Chao, Jeffrey A. Singer, Robert H. |
| description | Cellular messenger RNA levels are achieved by the combinatorial complexity of factors controlling transcription, yet the small number of molecules involved in these pathways fluctuates stochastically. It has not yet been experimentally possible to observe the activity of single polymerases on an endogenous gene to elucidate how these events occur in vivo. Here, we describe a method of fluctuation analysis of fluorescently labeled RNA to measure dynamics of nascent RNA—including initiation, elongation, and termination—at an active yeast locus. We find no transcriptional memory between initiation events, and elongation speed can vary by threefold throughout the cell cycle. By measuring the abundance and intranuclear mobility of an upstream transcription factor, we observe that the gene firing rate is directly determined by trans-activating factor search times. |
| doi_str_mv | 10.1126/science.1202142 |
| format | Article |
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It has not yet been experimentally possible to observe the activity of single polymerases on an endogenous gene to elucidate how these events occur in vivo. Here, we describe a method of fluctuation analysis of fluorescently labeled RNA to measure dynamics of nascent RNA—including initiation, elongation, and termination—at an active yeast locus. We find no transcriptional memory between initiation events, and elongation speed can vary by threefold throughout the cell cycle. By measuring the abundance and intranuclear mobility of an upstream transcription factor, we observe that the gene firing rate is directly determined by trans-activating factor search times.</description><identifier>ISSN: 0036-8075</identifier><identifier>EISSN: 1095-9203</identifier><identifier>DOI: 10.1126/science.1202142</identifier><identifier>PMID: 21512033</identifier><identifier>CODEN: SCIEAS</identifier><language>eng</language><publisher>Washington, DC: American Association for the Advancement of Science</publisher><subject>Adenosine Triphosphatases - genetics ; ATPases Associated with Diverse Cellular Activities ; Autocorrelation ; Biological and medical sciences ; Cell Cycle ; Cell nucleus ; Cell Nucleus - metabolism ; DNA Polymerase I - genetics ; Facilitated Diffusion ; Fluorescence ; Fundamental and applied biological sciences. Psychology ; Genes ; Genes, Fungal ; Glutamate Synthase - genetics ; Green Fluorescent Proteins ; Kinetics ; Microscopy, Fluorescence ; Models, Genetic ; Molecular and cellular biology ; Molecular genetics ; Molecules ; Promoter Regions, Genetic ; Real time ; Ribonucleic acid ; RNA ; RNA Polymerase II - metabolism ; RNA Precursors - genetics ; RNA Precursors - metabolism ; RNA, Fungal - biosynthesis ; RNA, Fungal - genetics ; RNA, Messenger - biosynthesis ; RNA, Messenger - genetics ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae - metabolism ; Saccharomyces cerevisiae Proteins - genetics ; Saccharomyces cerevisiae Proteins - metabolism ; Search time ; Spectrometry, Fluorescence ; Transcription Factors - metabolism ; Transcription, Genetic ; Transcription. Transcription factor. Splicing. Rna processing ; Transcriptional Activation ; Yeast ; Yeasts</subject><ispartof>Science (American Association for the Advancement of Science), 2011-04, Vol.332 (6028), p.475-478</ispartof><rights>Copyright © 2011 The American Association for the Advancement of Science</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011, American Association for the Advancement of Science</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c615t-681b44d71ae64e57f5756359ca04cd2f7e2c5a14ba02ddb92d1d752042372dc83</citedby><cites>FETCH-LOGICAL-c615t-681b44d71ae64e57f5756359ca04cd2f7e2c5a14ba02ddb92d1d752042372dc83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/29784132$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/29784132$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,776,780,799,881,2871,2872,27901,27902,57992,58225</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24153953$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21512033$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Larson, Daniel R.</creatorcontrib><creatorcontrib>Zenklusen, Daniel</creatorcontrib><creatorcontrib>Wu, Bin</creatorcontrib><creatorcontrib>Chao, Jeffrey A.</creatorcontrib><creatorcontrib>Singer, Robert H.</creatorcontrib><title>Real-Time Observation of Transcription Initiation and Elongation on an Endogenous Yeast Gene</title><title>Science (American Association for the Advancement of Science)</title><addtitle>Science</addtitle><description>Cellular messenger RNA levels are achieved by the combinatorial complexity of factors controlling transcription, yet the small number of molecules involved in these pathways fluctuates stochastically. 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| identifier | ISSN: 0036-8075 |
| ispartof | Science (American Association for the Advancement of Science), 2011-04, Vol.332 (6028), p.475-478 |
| issn | 0036-8075 1095-9203 |
| language | eng |
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| source | American Association for the Advancement of Science; Jstor Complete Legacy; MEDLINE |
| subjects | Adenosine Triphosphatases - genetics ATPases Associated with Diverse Cellular Activities Autocorrelation Biological and medical sciences Cell Cycle Cell nucleus Cell Nucleus - metabolism DNA Polymerase I - genetics Facilitated Diffusion Fluorescence Fundamental and applied biological sciences. Psychology Genes Genes, Fungal Glutamate Synthase - genetics Green Fluorescent Proteins Kinetics Microscopy, Fluorescence Models, Genetic Molecular and cellular biology Molecular genetics Molecules Promoter Regions, Genetic Real time Ribonucleic acid RNA RNA Polymerase II - metabolism RNA Precursors - genetics RNA Precursors - metabolism RNA, Fungal - biosynthesis RNA, Fungal - genetics RNA, Messenger - biosynthesis RNA, Messenger - genetics Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - metabolism Search time Spectrometry, Fluorescence Transcription Factors - metabolism Transcription, Genetic Transcription. Transcription factor. Splicing. Rna processing Transcriptional Activation Yeast Yeasts |
| title | Real-Time Observation of Transcription Initiation and Elongation on an Endogenous Yeast Gene |
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