Cell-by-Cell Dissection of Gene Expression and Chromosomal Interactions Reveals Consequences of Nuclear Reorganization 英文参考文献.docVIP

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Cell-by-Cell Dissection of Gene Expression and Chromosomal Interactions Reveals Consequences of Nuclear Reorganization 英文参考文献.doc

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Cell-by-Cell Dissection of Gene Expression and Chromosomal Interactions Reveals Consequences of Nuclear Reorganization 英文参考文献

o Openaccess,freelyavailableonline PL S BIOLOGY Cell-by-CellDissectionofGeneExpression andChromosomalInteractionsReveals ConsequencesofNuclearReorganization Brian Harmon, John Sedat* UniversityofCalifornia,SanFrancisco,California,UnitedStatesofAmerica The functional consequences of long-range nuclear reorganization were studied in a cell-by-cell analysis of gene expression and long-range chromosomal interactions in the Drosophila eye and eye imaginal disk. Position-effect variegationwasusedtostochasticallyperturbgeneexpressionandprobenuclearreorganization.Variegatinggenes onrearrangementsofChromosomesX,2,and3wereprobedforlong-rangeinteractionswithheterochromatin.Studies were conducted only in tissues known to express the variegating genes. Nuclear structure was revealed by fluorescence in situ hybridization with probes to the variegating gene and heterochromatin. Gene expression was determinedalternatelybyimmunofluorescence againstspecificproteinsandbyeyepigmentautofluorescence.This allowed cell-by-cell comparisons of nuclear architecture between cells in which the variegating gene was either expressed or silenced. Very strong correlations between heterochromatic association and silencing were found. Expressing cells showed a broad distribution of distances between variegating genes and their own centromeric heterochromatin,whilesilencedcellsshowedaverytightdistributioncenteredaroundveryshortdistances,consistent withinteractionbetweenthesilencedgenesandheterochromatin.Spatialandtemporalanalysisofinteractionswith heterochromatinindicatedthatvariegatinggenesprimarilyassociatewithheterochromatinincellsthathaveexited thecellcycle.Differentiationwasnotarequirementforassociation,andnodifferencesinassociationwereobserved betweencelltypes.Thus,long-rangeinteractionsbetweendistalchromosomeregionsandtheirownheterochromatin havefunctionalconsequencesfortheorganism. Citation:HarmonB,SedatJ(2005)Cell-by-celldissectionofgeneexpressionandchromosomalinteractionsrevealsconsequ