Synthetic Human β-Globin 5'HS2 Constructs Function as Partially Active Locus Control Regions.
EMBO Journal , Volume 12 p. 127- 134
Transgenes linked to the beta-globin locus control region (LCR) are transcribed in a copy-dependent manner that is independent of the integration site. It has previously been shown that the LCR 5'HS2 region does not require its NF-E2 dimer binding site for LCR activity. In this paper we analyse synthetic 5'HS2 core constructs containing point mutations in the other factor binding sites 3' of the NF-E2 dimer site. The results show that 5'HS2 core is a partially active LCR that functions in a concatamer of at least two copies but not when present as a single copy in transgenic mice and that no single binding site within 5'HS2 is required for position-independent expression. In addition, the H-BP factor is identical to upstream stimulatory factor (USF) and full enhancement levels by 5'HS2 core in MEL cells require a combination of all the factor binding sites. We suggest that 5'HS2 cores in a concatamer interact with each other to establish an area of open chromatin and that this process may be the basis of LCR function.
|*Genes, Regulator, *Genes, Synthetic, 0 (DNA-Binding Proteins), 0 (Transcription Factors), 125267-48-3 (erythroid-specific DNA-binding factor), 9004-22-2 (Globins), 9007-49-2 (dna), Animals, Base Sequence, Cell Line, DNA-Binding Proteins/genetics, DNA/genetics/isolation & purification, Fetus, Globins/*genetics, Human, Introns, Liver/physiology, Mice, Mice, Transgenic, Molecular Sequence Data, Mutagenesis, Site-Directed, Restriction Mapping, Support, Non-U.S. Gov't, Transcription Factors/genetics, Transfection, Zinc Fingers/genetics|
|Organisation||Erasmus MC: University Medical Center Rotterdam|
Ellis, J, Talbot, D, Dillon, N.O, & Grosveld, F.G. (1993). Synthetic Human β-Globin 5'HS2 Constructs Function as Partially Active Locus Control Regions. EMBO Journal, 12, 127–134. Retrieved from http://hdl.handle.net/1765/2483