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Echinobase
ECB-ART-36607
Proc Natl Acad Sci U S A 1997 Mar 18;946:2272-7. doi: 10.1073/pnas.94.6.2272.
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Enhancer blocking activity located near the 3'' end of the sea urchin early H2A histone gene.

Palla F , Melfi R , Anello L , Di Bernardo M , Spinelli G .


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The sea urchin early histone repeating unit contains one copy of each of the five histone genes whose coordinate expression during development is regulated by gene-specific elements. To learn how within the histone repeating unit a gene-specific activator can be prevented to communicate with the heterologous promoters, we searched for domain boundaries by using the enhancer blocking assay. We focused on the region near the 3'' end of the H2A gene where stage-specific nuclease cleavage sites appear upon silencing of the early histone genes. We demonstrated that a DNA fragment of 265 bp in length, defined as sns (for silencing nucleoprotein structure), blocked the enhancer activity of the H2A modulator in microinjected sea urchin embryos only when placed between the enhancer elements and the promoter. We also found that sns silenced the modulator elements even when placed at 2.7 kb from the promoter. By contrast, the enhancer activity of the modulator sequences, located downstream to the coding region, was not affected when sns was positioned in close proximity to the promoter. Finally, the H2A sns fragment placed between the simian virus 40 regulative region and the tk promoter repressed chloramphenicol acetyltransferase expression in transfected human cell lines. We conclude that 3'' end of the H2A gene contains sequence elements that behave as functional barriers of enhancer function in the enhancer blocking assay. Furthermore, our results also indicate that the enhancer blocking function of sns lacks enhancer and species specificity and that it can act in transient assays.

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Genes referenced: h2a LOC100887844 LOC100893907 LOC115917902 LOC115919910 LOC590297

References [+] :
Anello, Different micrococcal nuclease cleavage patterns characterize transcriptionally active and inactive sea-urchin histone genes. 1986, Pubmed, Echinobase