Journal article
Dysregulated human renin expression in transgenic mice carrying truncated genomic constructs: evidence supporting the presence of insulators at the renin locus
American Journal of Physiology - Renal Physiology, Vol.295(3), pp.F642-F653
07/16/2008
DOI: 10.1152/ajprenal.00384.2007
PMCID: PMC2536869
PMID: 18632798
Abstract
We previously generated transgenic mice carrying a large P1 artificial chromosome (PAC160) encompassing a 160-kb segment containing the human renin gene, two upstream genes, and one downstream gene. We also previously generated mutant PAC160 constructs lacking the distal enhancer and concluded it is required to maintain baseline expression of human renin, but is not required for tissue-specific, cell-specific, and regulated expression of renin in vivo. We now report two additional transgenic lines carrying random truncations of PAC160 upstream of the renin gene. Southern and PCR mapping studies indicate that the truncation break points in the two lines are located ∼10.4 and 2.5 kb upstream of the renin gene causing a deletion of all DNA upstream of the break. We tested the hypothesis that large-scale deletion of DNA upstream of the human renin gene including the enhancer would cause dysregulation of human renin expression. Phenotypically, these truncations cause a severe dysregulation of human renin expression, but remarkably, a preservation of the normal tissue-specific expression of the human ethanolamine kinase 2 (ETNK2) gene which lies immediately downstream of renin. Several functional binding sites for CTCF, a mammalian insulator protein, were identified in and around the renin and ETNK2 loci by gel shift and chromatin immunoprecipitation. We conclude that there are sequences in and around the renin and ETNK2 loci which act as boundaries between neighboring genes which insulate them from each other. The study illustrates the value of taking a much wider genomic perspective when studying mechanisms regulating gene expression.
Details
- Title: Subtitle
- Dysregulated human renin expression in transgenic mice carrying truncated genomic constructs: evidence supporting the presence of insulators at the renin locus
- Creators
- Xiyou Zhou - Roy J. and Lucille A. Carver College of MedicineEric T. Weatherford - University of IowaXuebo Liu - Molecular and Cellular Biology Graduate Program, Departments of Molecular Physiology and Biophysics and of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IowaElla Born - Molecular and Cellular Biology Graduate Program, Departments of Molecular Physiology and Biophysics and of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IowaHenry L. Keen - University of IowaCurt D. Sigmund - University of Iowa
- Resource Type
- Journal article
- Publication Details
- American Journal of Physiology - Renal Physiology, Vol.295(3), pp.F642-F653
- Publisher
- American Physiological Society
- DOI
- 10.1152/ajprenal.00384.2007
- PMID
- 18632798
- PMCID
- PMC2536869
- ISSN
- 0363-6127
- eISSN
- 1522-1466
- Language
- English
- Date published
- 07/16/2008
- Academic Unit
- Molecular Physiology and Biophysics; Neuroscience and Pharmacology; Endocrinology and Metabolism; Internal Medicine; Iowa Institute of Human Genetics
- Record Identifier
- 9984359850002771
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