PuSH - Publication Server of Helmholtz Zentrum München

Meggendorfer, M. ; Weierich, C.* ; Wolff, H. ; Brack-Werner, R. ; Cremer, T.*

Functional nuclear topography of transcriptionally inducible extra-chromosomal transgene clusters.

Chromosome Res. 18, 401-417 (2010)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
A new experimental approach was designed to test different predictions of current models of the nuclear architecture with respect to the topography of transcription. We constructed a plasmid, termed pIndi, which carries a reporter gene coding for a red cytoplasmic fluorescent reporter protein. Transcription of the reporter gene is regulated by the inducible promoter of the human immunodeficiency virus (HIV) and is strongly dependent on the HIV-1 Tat protein. Expressing the red fluorescent reporter protein allowed us to distinguish between cells with active and silent reporter genes. Importantly, transient transfection resulted in the clustering of plasmids, forming one or several extra-chromosomal pIndi bodies. Repetitive lac operator sequences in pIndi allowed us to visualize these bodies in living cells by the binding of LacI proteins tagged with a fluorescent protein. Using this model, we analyzed the three-dimensional nuclear topography of pIndi bodies with active or silent reporter genes. Our results are compatible with predictions of the chromosome territory-interchromatin compartment (CT-IC) model. We demonstrate that pIndi bodies localize in the IC, both in the silent and active state. Activation of transgene transcription resulted in the recruitment of RNA polymerase II and NFkappaB and a closer positioning to splicing speckles.
Altmetric
Additional Metrics?
[➜Log in]
Edit extra informations Login
Publication type Article: Journal article
Document type Scientific Article
Keywords Chromosome territory; Interchromatin compartment; Nuclear organization; Models of nuclear architecture
ISSN (print) / ISBN 0967-3849
e-ISSN 1573-6849
Journal Chromosome Research
Quellenangaben Volume: 18, Issue: 4, Pages: 401-417 Article Number: , Supplement: ,
Publisher Springer
Reviewing status Peer reviewed
Institute(s) Institute of Virology (VIRO)