Effects of methylation of non-CpG sequence in the promoter region on the expression of human synaptotagmin XI (syt11).
We have studied the effects of methylation of the promoter region on the expression of human synaptotagmin XI (syt11), a gene implicated in the onset of schizophrenia. Sequence analysis showed that cytosine residues not in the CpG sequence, but still within the promoter region of the gene, are partially methylated. The methylated cytosine residues are located in the mRNA-coding (minus) strand of the promoter region (mCmCTTmCTTmCmC). Gel mobility shift assays showed that when the cytosine residues are methylated, the binding activity of an Sp family protein, a transcription factor, to the region is significantly reduced. Furthermore, transient transcription assays using artificially methylated promoter sequences showed that methylation did reduce the expression of the reporter gene. The biological significance of the finding is discussed in respect to the effect of methylation of non-CpG sequences in promoter regions on gene expression.
Inoue
Shinichi
S
Kazusa Research Institute, 2-6-7, Kazusa-kamatari, Kisarazu, Chiba 292-0818, Japan. sinoue@kazusa.or.jp
Oishi
Michio
M
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D001667:Binding, Competitive; D016466:CHO Cells; D002135:Calcium-Binding Proteins / Q000235:genetics*; D002460:Cell Line; D045744:Cell Line, Tumor; D018899:CpG Islands / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D003596:Cytosine / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D016329:Sp1 Transcription Factor / Q000378:metabolism; D051705:Sp3 Transcription Factor; D013447:Sulfites / Q000737:chemistry; D050857:Synaptotagmins; D014157:Transcription Factors / Q000378:metabolism; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection
D002135:Calcium-Binding Proteins; D004268:DNA-Binding Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D009841:Oligonucleotides; C494265:SYT11 protein, human; D016329:Sp1 Transcription Factor; D013447:Sulfites; D014157:Transcription Factors; D050857:Synaptotagmins; D051705:Sp3 Transcription Factor; D003596:Cytosine; D004247:DNA; D008156:Luciferases; C042345:hydrogen sulfite