The activity of the scs and scs' insulator elements is not dependent on chromosomal context.

Molecular and Cellular Biology
M DunawayH L Yuen

Abstract

Sequence elements that protect a reporter gene from chromosomal position effects or that block enhancer-activated transcription are called insulators. Using a plasmid-based microinjection assay with Xenopus laevis oocytes, we show that the heterologous Drosophila melanogaster scs and scs' insulator elements do not require chromosomal context to block enhancer-activated transcription. A single insulator element partially blocks enhancer-activated transcription, indicating that each element operates independently rather than as part of a pair. Deletion analysis of the 1.8-kb scs element identified a 220-bp fragment from one of the DNase I-hypersensitive regions that has full blocking activity in the oocyte assay. This fragment corresponds to the critical region of the scs mapped in previous studies with Drosophila. A time course of transcription shows that the scs blocks enhancer-activated transcription as early as transcription can be detected, about 30 min after injection. Complete assembly of the DNA template into nucleosomes requires 4 h. The scs and scs' sequences do not block site-specific recombination by FLP recombinase, implying that insulators do not operate by a general mechanism that physically sequesters the DNA. The...Continue Reading

References

May 1, 1992·Molecular and Cellular Biology·R Kellum, P Schedl
Dec 20, 1985·Nucleic Acids Research·P Labhart, R H Reeder
Aug 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·D ShoreR L Baldwin
Jan 1, 1984·Cold Spring Harbor Symposia on Quantitative Biology·M McLeodJ Broach

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Citations

Dec 23, 2009·Developmental Biology·Michael Bulger, Mark Groudine
May 14, 1999·Current Opinion in Genetics & Development·A C Bell, G Felsenfeld
Nov 9, 2001·Annual Review of Genetics·T I Gerasimova, V G Corces
Nov 6, 2008·Molecular Therapy : the Journal of the American Society of Gene Therapy·Trine DalsgaardJacob G Mikkelsen
Dec 13, 2000·Proceedings of the National Academy of Sciences of the United States of America·W Wei, M D Brennan
Jan 15, 2005·The EMBO Journal·Stefan Ludwig AmeresChristian Berens
Jan 30, 2004·Molecular and Cellular Biology·Emily J KuhnPamela K Geyer
Oct 25, 2000·Molecular and Cellular Biology·O AlbagliF Puvion-Dutilleul
Mar 25, 2005·Bioorganicheskaia khimiia·A V SassE D Sverdlov

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