title: Uniform distribution of elongating RNA polymerase II complexes in transcribed gene locus.
abstract: The intensity of gene transcription is generally reflected by the level of RNA polymerase II (RNAPII) recruitment to the gene. However, genome-wide studies of polymerase occupancy indicate that RNAPII distribution varies among genes. In some loci more polymerases are found in the 5' region, whereas in other loci, in the 3' region of the gene. We studied the distribution of elongating RNAPII complexes at highly transcribed GAL-VPS13 locus in Saccharomyces cerevisiae and found that in the cell population the amount of polymerases gradually decreased toward the 3' end of the gene. However, the conventional chromatin immunoprecipitation assay averages the signal from the cell population, and no data on single cell level can be gathered. To study the spacing of elongating polymerases on single chromosomes, we used a sequential chromatin immunoprecipitation assay for the detection of multiple RNAPII complexes on the same DNA fragment. Our results demonstrate uniform distribution of elongating polymerases throughout all regions of the GAL-VPS13 gene.
authors: Institute of Molecular and Cell Biology, University of Tartu, Riia 23, Tartu 51010, Estonia.
authors: Kadri
authors: K
authors: Peil
authors: Signe
authors: S
authors: Värv
authors: Marko
authors: M
authors: Lõoke
authors: Kersti
authors: K
authors: Kristjuhan
authors: Arnold
authors: A
authors: Kristjuhan
references: Cell. 2005 Jun 17;121(6):913-23
references: 15960978
references: Nat Genet. 2004 Aug;36(8):900-5
references: 15247917
references: Curr Biol. 2009 Dec 15;19(23):2037-42
references: 19931455
references: Proc Natl Acad Sci U S A. 2010 Jun 22;107(25):11325-30
references: 20534568
references: Science. 2011 Apr 22;332(6028):475-8
references: 21512033
references: Cell. 2003 Dec 12;115(6):751-63
references: 14675539
references: Science. 2009 Jul 31;325(5940):626-8
references: 19644123
references: Mol Cell. 2006 Nov 3;24(3):469-79
references: 17081995
references: PLoS Biol. 2006 Oct;4(10):e309
references: 17048983
references: Mol Cell. 2010 Apr 23;38(2):202-10
references: 20417599
references: EMBO J. 2004 Oct 27;23(21):4243-52
references: 15457216
references: Nat Struct Mol Biol. 2010 Jun;17(6):745-52
references: 20453861
references: Genome Biol. 2010;11(7):R75
references: 20637075
references: Curr Biol. 2006 May 23;16(10):1018-25
references: 16713960
references: Mol Cell. 2002 Oct;10(4):925-33
references: 12419235
references: Nature. 2002 Feb 21;415(6874):929-33
references: 11859374
references: Nat Struct Mol Biol. 2008 Jan;15(1):71-8
references: 18157150
references: Nat Struct Mol Biol. 2008 Dec;15(12):1263-71
references: 19011635
references: Science. 2008 Jun 6;320(5881):1344-9
references: 18451266
references: Proc Natl Acad Sci U S A. 1996 May 28;93(11):5208-12
references: 8643554
references: Mol Cell. 2006 Dec 8;24(5):735-746
references: 17157256
references: Cell. 1998 Nov 25;95(5):717-28
references: 9845373
references: PLoS One. 2010 Nov 16;5(11):e15442
references: 21103382
references: Biochem Biophys Res Commun. 2007 Jun 29;358(2):666-71
references: 17498649
references: Nucleic Acids Res. 2010 Aug;38(14):4651-64
references: 20385590
references: Nature. 2011 Jan 20;469(7330):368-73
references: 21248844
references: Mol Cell. 2005 Apr 1;18(1):97-108
references: 15808512
references: Mol Cell Biol. 2004 Dec;24(23):10111-7
references: 15542822