We have used a kinetic analysis to distinguish possible mechanisms of activation of transcription of the different genes in the human beta globin locus. Based on in situ studies at the single-cell level we have previously suggested a dynamic mechanism of single genes alternately interacting with the locus control region (LCR) to activate transcription. However, those steady-state experiments did not allow a direct measurement of the dynamics of the mechanism and the presence of loci with in situ primary transcript signals from two beta-like genes in cis has left open the possibility that multiple genes in the locus could initiate transcription simultaneously. Kinetic assays involving removal of a block to transcription elongation in conjunction with RNA FISH show that multiple beta gene primary transcript signals in cis represent a transition between alternating transcriptional periods of single genes, supporting a dynamic interaction mechanism.

Animals, Base Sequence, Binding, Competitive/genetics, Cells, Cultured, Chromatin/*genetics/metabolism, Dactinomycin/pharmacology, Dichlororibofuranosylbenzimidazole/pharmacology, Fetus, Globins/genetics/metabolism, Humans, In Situ Hybridization, Fluorescence, Kinetics, Liver, Locus Control Region/drug effects, Mice, Mice, Transgenic, Molecular Sequence Data, Nucleic Acid Synthesis Inhibitors/pharmacology, Research Support, Non-U.S. Gov't, Transcription, Genetic/drug effects/*physiology
dx.doi.org/10.1093/emboj/17.20.6020, hdl.handle.net/1765/12805
EMBO Journal
Erasmus MC: University Medical Center Rotterdam

Gribnau, J.H, de Boer, E, Trimborn, T, Wijgerde, M.G.J.M, Milot, E, Fraser, P.J, & Grosveld, F.G. (1998). Chromatin interaction mechanism of transcriptional control in vivo. EMBO Journal, 17(20), 6020–6027. doi:10.1093/emboj/17.20.6020