5-Azacytidine-induced reactivation of the human X chromosome-linked PGK1 gene is associated with a large region of cytosine demethylation in the 5 prime CpG island
Journal Article
·
· Proceedings of the National Academy of Sciences of the United States of America; (United States)
- Univ. of Washington, Seattle (USA)
Hamster-human cell hybrids containing an inactive human X chromosome were treated with 5-azacytidine and derived clones were examined for phosphoglycerate kinase activity and cytosine methylation in the human PGK1 (X chromosome-linked phosphoglycerate kinase) gene. Comparisons between expressing and nonexpressing clones indicated that demethylation of several methylation-sensitive restriction sites outside of the 5{prime} CpG island were unnecessary for expression. High-resolution polyacrylamide gel analysis of 25 Hpa II, Hha I, and Tha I sites revealed that all clones expressing PGK1 were unmethylated in a large region of the CpG island that includes the transcription start site and 400 base pairs upstream. Many nonexpressing clones had discontinuous patterns of demethylation. Remethylation was often observed in subclones of nonexpressing hybrids. These data suggest that a specific zone of methylation-free DNA within the PGK1 promoter is required for transcription. In addition, the presence of neighboring methylcytosines appears to decrease the heritable stability of unmethylated CpGs in this region.
- OSTI ID:
- 5393219
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America; (United States), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (United States) Vol. 87:11; ISSN 0027-8424; ISSN PNASA
- Country of Publication:
- United States
- Language:
- English
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Wed Oct 31 23:00:00 EST 1990
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Fri Feb 14 23:00:00 EST 1992
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·
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Related Subjects
550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMALS
ANTIMETABOLITES
AZINES
BIOLOGICAL EFFECTS
CHEMICAL REACTIONS
CHROMOSOMES
CYTOSINE
DNA
DNA HYBRIDIZATION
DNA METHYLASES
DRUGS
ENZYMES
HETEROCHROMOSOMES
HETEROCYCLIC COMPOUNDS
HUMAN X CHROMOSOME
HYBRIDIZATION
INACTIVATION
LYASES
MAMMALS
MAN
METHYLATION
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PRIMATES
PROTEINS
PYRIMIDINES
VERTEBRATES
X CHROMOSOME
59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMALS
ANTIMETABOLITES
AZINES
BIOLOGICAL EFFECTS
CHEMICAL REACTIONS
CHROMOSOMES
CYTOSINE
DNA
DNA HYBRIDIZATION
DNA METHYLASES
DRUGS
ENZYMES
HETEROCHROMOSOMES
HETEROCYCLIC COMPOUNDS
HUMAN X CHROMOSOME
HYBRIDIZATION
INACTIVATION
LYASES
MAMMALS
MAN
METHYLATION
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PRIMATES
PROTEINS
PYRIMIDINES
VERTEBRATES
X CHROMOSOME