Long-range functional interactions between genes and regulatory elements
The mouse b-globin locus contains four genes that are expressed throughout development in the same order they are arranged along chromosome 7. Upstream of the genes is a powerful regulatory element called the locus control region (LCR). The LCR is absolutely required for high-level expression of the...
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ndltd-bl.uk-oai-ethos.bl.uk-5973292015-03-20T06:08:52ZLong-range functional interactions between genes and regulatory elementsCarter, D. P. F.2004The mouse b-globin locus contains four genes that are expressed throughout development in the same order they are arranged along chromosome 7. Upstream of the genes is a powerful regulatory element called the locus control region (LCR). The LCR is absolutely required for high-level expression of the b-globin genes. The models to explain how LCRs (and enhancers in general) are able to activate expression of the genes fall into two categories: contact and non-contact. The contact models propose that the LCR and the gene form a physical interaction that leads to active transcription. The non-contact models propose that the LCR alters the chromatin topology or nuclear organisation of the locus, or acts as a nucleation point for factors that polymerise along the chromatin fibre to activate the gene. A contact model has been inferred from indirect observations, <i>in vitro</i> experiments, and oversimplified assumptions. In the absence of a physical assay that is able to determine the three-dimensional arrangements of the b-globin locus the mechanisms of LCR activation remains open to speculation. In this thesis I describe the development of a novel technique called RNA tagging and recovery of associated proteins (RNA TRAP). RNA TRAP uses the power of RNA FISH to target an enzyme to the site of a specific actively transcribing gene. The enzyme catalayses the deposition of a biotin tag onto the chromatin in the vicinity of transcription, which acts as a handle to isolate and identify chromatin elements in proximity to the actively transcribed gene. Using RNA TRAP I have shown that elements of the LCR are in significantly physical proximity to the genes they activate <i>in vivo.</i> This is the first physical demonstration of the interaction of the LCR with a gene, and gives insight into the regulation of genes by enhancers.591.35University of Cambridgehttp://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.597329Electronic Thesis or Dissertation |
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591.35 Carter, D. P. F. Long-range functional interactions between genes and regulatory elements |
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The mouse b-globin locus contains four genes that are expressed throughout development in the same order they are arranged along chromosome 7. Upstream of the genes is a powerful regulatory element called the locus control region (LCR). The LCR is absolutely required for high-level expression of the b-globin genes. The models to explain how LCRs (and enhancers in general) are able to activate expression of the genes fall into two categories: contact and non-contact. The contact models propose that the LCR and the gene form a physical interaction that leads to active transcription. The non-contact models propose that the LCR alters the chromatin topology or nuclear organisation of the locus, or acts as a nucleation point for factors that polymerise along the chromatin fibre to activate the gene. A contact model has been inferred from indirect observations, <i>in vitro</i> experiments, and oversimplified assumptions. In the absence of a physical assay that is able to determine the three-dimensional arrangements of the b-globin locus the mechanisms of LCR activation remains open to speculation. In this thesis I describe the development of a novel technique called RNA tagging and recovery of associated proteins (RNA TRAP). RNA TRAP uses the power of RNA FISH to target an enzyme to the site of a specific actively transcribing gene. The enzyme catalayses the deposition of a biotin tag onto the chromatin in the vicinity of transcription, which acts as a handle to isolate and identify chromatin elements in proximity to the actively transcribed gene. Using RNA TRAP I have shown that elements of the LCR are in significantly physical proximity to the genes they activate <i>in vivo.</i> This is the first physical demonstration of the interaction of the LCR with a gene, and gives insight into the regulation of genes by enhancers. |
author |
Carter, D. P. F. |
author_facet |
Carter, D. P. F. |
author_sort |
Carter, D. P. F. |
title |
Long-range functional interactions between genes and regulatory elements |
title_short |
Long-range functional interactions between genes and regulatory elements |
title_full |
Long-range functional interactions between genes and regulatory elements |
title_fullStr |
Long-range functional interactions between genes and regulatory elements |
title_full_unstemmed |
Long-range functional interactions between genes and regulatory elements |
title_sort |
long-range functional interactions between genes and regulatory elements |
publisher |
University of Cambridge |
publishDate |
2004 |
url |
http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.597329 |
work_keys_str_mv |
AT carterdpf longrangefunctionalinteractionsbetweengenesandregulatoryelements |
_version_ |
1716795451274952704 |