Using molecular tethering to analyze the role of nuclear compartmentalization in the regulation of mammalian gene activity.
The mammalian nucleus has a complex structural organization that dynamically interacts with the genome. Chromatin is organized into discrete domains by association with distinct nuclear compartments enriched in structural and regulatory proteins. Growing evidence suggests that gene activity is modulated by interactions with these sub-nuclear compartments. Therefore, analyzing how nuclear architecture controls genome activity will be necessary to fully understand complex biological processes such as development and disease. In this article we describe a molecular methodology involving inducible tethering that can be used to position genes at the inner nuclear membrane (INM)-lamina compartment. The consequences of such directed re-positioning on gene activity or other DNA transactions can then be analyzed. This approach can be generalized and extended to position genes or chromosomal domains within other nuclear compartments thereby greatly facilitating the analysis of nuclear structure and its impact on genome activity.
Reddy
K L
KL
Department of Molecular Genetics and Cell Biology, The University of Chicago, Howard Hughes Medical Institute, GCIS W522, 929 E 57th Street, Chicago, IL 60637, USA.
Singh
H
H
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly / Q000502:physiology; D045584:Chromosome Positioning; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D018274:Electroporation; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007763:Lac Operon; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008322:Mammals / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods*; D015530:Nuclear Matrix / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D015341:Nucleic Acid Probes / Q000032:analysis / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D013937:Thymidine Kinase / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology
D001426:Bacterial Proteins; D002843:Chromatin; C485182:Cyan Fluorescent Protein; D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; D015341:Nucleic Acid Probes; D011993:Recombinant Fusion Proteins; C092710:emerin; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins; D013937:Thymidine Kinase