Galactose repressor mediated intersegmental chromosomal connections in Escherichia coli.
By microscopic analysis of fluorescent-labeled GalR, a regulon-specific transcription factor in Escherichia coli, we observed that GalR is present in the cell as aggregates (one to three fluorescent foci per cell) in nongrowing cells. To investigate whether these foci represent GalR-mediated association of some of the GalR specific DNA binding sites (gal operators), we used the chromosome conformation capture (3C) method in vivo. Our 3C data demonstrate that, in stationary phase cells, many of the operators distributed around the chromosome are interacted. By the use of atomic force microscopy, we showed that the observed remote chromosomal interconnections occur by direct interactions between DNA-bound GalR not involving any other factors. Mini plasmid DNA circles with three or five operators positioned at defined loci showed GalR-dependent loops of expected sizes of the intervening DNA segments. Our findings provide unique evidence that a transcription factor participates in organizing the chromosome in a three-dimensional structure. We believe that these chromosomal connections increase local concentration of GalR for coordinating the regulation of widely separated target genes, and organize the chromosome structure in space, thereby likely contributing to chromosome compaction.
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Zhong
Z
Qian
Emilios K
EK
Dimitriadis
Rotem
R
Edgar
Prahathees
P
Eswaramoorthy
Sankar
S
Adhya
D001665:Binding Sites; D002875:Chromosomes / Q000378:metabolism*; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism*; D005690:Galactose / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D018625:Microscopy, Atomic Force / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C050993:Galactose repressor proteins; D011956:Receptors, Cell Surface; D012097:Repressor Proteins; D014157:Transcription Factors; C039707:galactose receptor; D004247:DNA; D005690:Galactose