Chicken vigilin gene: a distinctive pattern of hypersensitive sites is characteristic for its transcriptional activity

Vigilin, a multidomain hn-ribonucleo-K-homologous protein, is part of a ribonucleoprotein complex with cognate tRNA and is found in both the nucleus and the cytoplasm. In an approach to identify genomic regions involved in regulation of the chicken vigilin gene, we carried out transfection studies w...

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Bibliographic Details
Published inBiochemical journal Vol. 326 ( Pt 2); no. 2; pp. 601 - 607
Main Authors Grünweller, A, Purschke, W G, Kügler, S, Kruse, C, Müller, P K
Format Journal Article
LanguageEnglish
Published England 01.09.1997
Subjects
Animals
Base Sequence
Carrier Proteins
Chick Embryo
Chickens
Deoxyribonuclease I - genetics
DNA Fragmentation
Electrophoresis, Polyacrylamide Gel
Fibroblasts - enzymology
Fibroblasts - metabolism
Gene Expression Regulation, Developmental
Lymphocytes - enzymology
Molecular Sequence Data
Promoter Regions, Genetic
Protein Biosynthesis
Proteins - genetics
Regulatory Sequences, Nucleic Acid
RNA-Binding Proteins
Tendons - cytology
Tendons - enzymology
Tendons - metabolism
Transcription, Genetic
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Summary:Vigilin, a multidomain hn-ribonucleo-K-homologous protein, is part of a ribonucleoprotein complex with cognate tRNA and is found in both the nucleus and the cytoplasm. In an approach to identify genomic regions involved in regulation of the chicken vigilin gene, we carried out transfection studies with a reporter gene in suitable chicken cells. After including a distantly positioned 5'-sequence in the construct, we observed a 10.5-fold increase in luciferase (EC 1. 13.12.7) expression compared with basal promoter activity. Accordingly, chromatin analysis of freshly isolated embryonic tendon fibroblasts with high levels of vigilin mRNA expression shows a DNase-I-hypersensitive site (DHS1) localized 2.2 kb upstream of the transcriptional start site. Similarly, phytohaemagglutinin-stimulated lymphocytes with a 4-fold elevated expression of vigilin mRNA compared with resting lymphocytes also exhibited this unique DHS, having switched from that found at 3.3 kb (DHS2) in resting lymphocytes. Furthermore, using gel-retardation experiments with DNA representing either DHS1 or DHS2, a specific interaction with chicken nuclear extracts was seen.
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ISSN:0264-6021
1470-8728
DOI:10.1042/bj3260601