Genomic organization and regulatory elements of the rat latexin gene, which is expressed in a cell type-specific manner in both central and peripheral nervous systems.
Latexin, a carboxypeptidase A inhibitor, is expressed in a cell type-specific manner in both central and peripheral nervous systems in the rat. In the neocortex, a specific subpopulation of neurons in layers V and VI expresses latexin. In the primary sensory ganglia, the expression is restricted to smaller diameter neurons. As a first step to clarify regulatory mechanisms underlying cell type-specific expression of latexin, we have determined the organization of the rat latexin gene and analyzed its regulatory elements. The latexin gene spans approximately 5.8 kb, and consists of six exons and five introns. Three transcription initiation sites were mapped. The upstream region lacks typical TATA or CAAT boxes but has several GC-rich sites. To assess promoter activity, the luciferase reporter gene fused to the 5'-flanking region (6.4 kb) of the latexin gene was transiently transfected into several cell lines. Luciferase activity was 2-8 times higher in latexin-expressing cells (PC12) than non-expressing cells (NS20 and L6). Deletion analysis with PC12 cells revealed that a core promoter is located between nucleotide positions -261 and -201 relative to the A of the initiation codon. Nerve growth factor (NGF)-responsive element(s) is located between positions -518 and -262, in which AP-1, AP-2 and NF-kappaB binding sites are found. Furthermore, we demonstrate that a 1.3 kb genomic fragment containing the first intron has transcriptional enhancing activity in PC12 cells. These results suggest that up and downstream regulatory elements are involved in the control of cell type-specific expression of latexin.
Miyasaka
N
N
Mitsubishi Kasei Institute of Life Sciences, 11 Minamiooya, Machida-shi, Tokyo 194-8511, Japan. miyasaka@libra.ls.m-kagaku.co.jp
Hatanaka
Y
Y
Jin
M
M
Arimatsu
Y
Y
D020121:5' Untranslated Regions / Q000502:physiology; D000818:Animals; D000941:Antigens / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002268:Carboxypeptidases / Q000032:analysis; D043422:Carboxypeptidases A; D002540:Cerebral Cortex / Q000737:chemistry / Q000166:cytology; D003001:Cloning, Molecular; D017931:DNA Primers; D004742:Enhancer Elements, Genetic / Q000502:physiology; D017950:Ganglia, Sensory / Q000737:chemistry / Q000166:cytology; D005786:Gene Expression Regulation / Q000502:physiology; D017930:Genes, Reporter; D016678:Genome; D007438:Introns / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009475:Neurons, Afferent / Q000737:chemistry* / Q000201:enzymology; D016716:PC12 Cells; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection
D020121:5' Untranslated Regions; D000941:Antigens; D017931:DNA Primers; C086812:Lxn protein, rat; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D008156:Luciferases; D002268:Carboxypeptidases; D043422:Carboxypeptidases A