title: DNA methylation induced changes in chromatin conformation of the promoter of the vitellogenin II gene of Japanese quail during aging.
abstract: One approach to the understanding of the molecular basis of aging in higher organisms may be to use genes whose timing and rate of expression during the life span run parallel with specific functions that can be monitored. The genes for egg proteins, such as vitellogenin (VTG), which is expressed in the liver, and ovalbumin, lysozyme etc. that are expressed in the oviduct of birds, meet these requirements. Egg laying function is dependent on the production of these proteins, which, in turn, depends on the expression of their genes. In this communication we present the age-related studies on the VTG II gene of the bird, Japanese quail. The gene is expressed only in the liver and its expression is considerably lower in old birds that do not lay eggs. Comparison of the promoter region of the gene carrying the two important cis-acting elements, estrogen responsive element (ERE) and progesterone responsive element (PRE), shows it to be 100% homologous to the corresponding region of the chicken VTG II gene. Methylation of DNA and conformation of chromatin of this region were studied, as they are known to be important for regulation of expression of genes. Our studies show that in the liver of adult female quails which lay eggs, a -CCGG- sequence located in this region is hypomethylated, and the chromatin encompassing this region of the gene is relaxed. In the old, the -CCGG- sequence is hypermethylated and the chromatin is compact. This is correlated with a decrease in the expression of the gene and decrease in egg production. Further, electrophoretic mobility shift assay (EMSA) shows that the levels/affinity of specific trans-acting factors that bind to ERE and PRE present in the region, are not different in adult and old birds. Hence the methylation status of the -CCGG- sequence that is located in-between the ERE and the PRE may be crucial for the conformation of chromatin and availability of these two important cis-acting elements for the binding of the trans-acting factors. This, in turn, may downregulate the expression of the gene in old birds.
authors.affiliation: Molecular Biology Laboratory, Department of Zoology, Banaras Hindu University, Varanasi, India.
authors.forename: Sanjay
authors.initials: S
authors.lastname: Gupta
authors.forename: Rashmi U
authors.initials: RU
authors.lastname: Pathak
authors.forename: Madhu S
authors.initials: MS
authors.lastname: Kanungo
mesh_terms: D000375:Aging / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002843:Chromatin / Q000235:genetics; D003370:Coturnix / Q000235:genetics; D019175:DNA Methylation; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D003850:Deoxyribonuclease I; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D008836:Micrococcal Nuclease; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014819:Vitellogenins / Q000235:genetics*
chemical_list: D002843:Chromatin; D018076:DNA, Complementary; D014819:Vitellogenins; D003850:Deoxyribonuclease I; D008836:Micrococcal Nuclease