title: Determinants of Chromosome Architecture: Insulator Pairing in cis and in trans.
abstract: The chromosomes of multicellular animals are organized into a series of topologically independent looped domains. This domain organization is critical for the proper utilization and propagation of the genetic information encoded by the chromosome. A special set of architectural elements, called boundaries or insulators, are responsible both for subdividing the chromatin into discrete domains and for determining the topological organization of these domains. Central to the architectural functions of insulators are homologous and heterologous insulator:insulator pairing interactions. The former (pairing between copies of the same insulator) dictates the process of homolog alignment and pairing in trans, while the latter (pairing between different insulators) defines the topology of looped domains in cis. To elucidate the principles governing these architectural functions, we use two insulators, Homie and Nhomie, that flank the Drosophila even skipped locus. We show that homologous insulator interactions in trans, between Homie on one homolog and Homie on the other, or between Nhomie on one homolog and Nhomie on the other, mediate transvection. Critically, these homologous insulator:insulator interactions are orientation-dependent. Consistent with a role in the alignment and pairing of homologs, self-pairing in trans is head-to-head. Head-to-head self-interactions in cis have been reported for other fly insulators, suggesting that this is a general principle of self-pairing. Homie and Nhomie not only pair with themselves, but with each other. Heterologous Homie-Nhomie interactions occur in cis, and we show that they serve to delimit a looped chromosomal domain that contains the even skipped transcription unit and its associated enhancers. The topology of this loop is defined by the heterologous pairing properties of Homie and Nhomie. Instead of being head-to-head, which would generate a circular loop, Homie-Nhomie pairing is head-to-tail. Head-to-tail pairing in cis generates a stem-loop, a configuration much like that observed in classical lampbrush chromosomes. These pairing principles provide a mechanistic underpinning for the observed topologies within and between chromosomes.
authors.affiliation: Deptartment of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of America.
authors.forename: Miki
authors.initials: M
authors.lastname: Fujioka
authors.affiliation: Departments of Biology and Biochemistry, Widener University, Chester, Pennsylvania, United States of America.
authors.forename: Hemlata
authors.initials: H
authors.lastname: Mistry
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, New Jersey, United States of America.|Laboratory of Gene Expression Regulation in Development, Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Paul
authors.initials: P
authors.lastname: Schedl
authors.affiliation: Deptartment of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of America.
authors.forename: James B
authors.initials: JB
authors.lastname: Jaynes
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D059006:Chromosomes, Insect / Q000737:chemistry* / Q000502:physiology; D029721:Drosophila Proteins / Q000235:genetics*; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000502:physiology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics*; D038101:Insulator Elements / Q000235:genetics*; D007763:Lac Operon; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D014157:Transcription Factors; C052837:eve protein, Drosophila; D049452:Green Fluorescent Proteins