Publikationsdatum:
2013-10-05
Beschreibung:
The chromatin immunoprecipitation (ChIP) assay is widely used to capture interactions between chromatin and regulatory proteins, but it is unknown how stable most native interactions are. Although live-cell imaging suggests short-lived interactions at tandem gene arrays, current methods cannot measure rapid binding dynamics at single-copy genes. We show, by using a modified ChIP assay with subsecond temporal resolution, that the time dependence of formaldehyde cross-linking can be used to extract in vivo on and off rates for site-specific chromatin interactions varying over a ~100-fold dynamic range. By using the method, we show that a regulatory process can shift weakly bound TATA-binding protein to stable promoter interactions, thereby facilitating transcription complex formation. This assay provides an approach for systematic, quantitative analyses of chromatin binding dynamics in vivo.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3997053/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3997053/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Poorey, Kunal -- Viswanathan, Ramya -- Carver, Melissa N -- Karpova, Tatiana S -- Cirimotich, Shana M -- McNally, James G -- Bekiranov, Stefan -- Auble, David T -- GM55763/GM/NIGMS NIH HHS/ -- R01 GM055763/GM/NIGMS NIH HHS/ -- Intramural NIH HHS/ -- New York, N.Y. -- Science. 2013 Oct 18;342(6156):369-72. doi: 10.1126/science.1242369. Epub 2013 Oct 3.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Biochemistry and Molecular Genetics, University of Virginia Health System, Charlottesville, VA 22908, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/24091704" target="_blank"〉PubMed〈/a〉
Schlagwort(e):
Adenosine Triphosphatases/chemistry/metabolism
;
Chromatin/chemistry/*metabolism
;
Chromatin Immunoprecipitation/*methods
;
Cross-Linking Reagents/chemistry
;
DNA-Binding Proteins/chemistry/metabolism
;
Formaldehyde/chemistry
;
Gene Dosage
;
*Gene Expression Regulation
;
Kinetics
;
Promoter Regions, Genetic
;
Saccharomyces cerevisiae Proteins/chemistry/metabolism
;
TATA-Binding Protein Associated Factors/chemistry/metabolism
;
TATA-Box Binding Protein/chemistry/*metabolism
;
Transcription Factors/chemistry/metabolism
Print ISSN:
0036-8075
Digitale ISSN:
1095-9203
Thema:
Biologie
,
Chemie und Pharmazie
,
Informatik
,
Medizin
,
Allgemeine Naturwissenschaft
,
Physik
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