Regional methylation of the 5' end CpG island of BRCA1 is associated with reduced gene expression in human somatic cells.
In mammalians, demethylation of specific promoter regions often correlates with gene activation; inversely, dense methylation of CpG islands leads to gene silencing, probably mediated by methyl-CpG binding proteins. In cell lines and cancers, inhibition of tissue-specific genes and tumor suppressor genes expression seems to be related to such hypermethylation. The 5' end of the breast cancer predisposition gene BRCA1 is embedded in a large CpG island of approximately 2.7 kb in length. In human sporadic breast cancers, the down-regulation of BRCA1 does not seem to be related to BRCA1 gene alterations. Southern blot analysis and the bisulfite sequencing method indicate that the BRCA1 CpG island is regionally methylated in all human tissues analyzed and unmethylated in the gametes, suggesting a role for DNA methylation in the control of gene expression. We have therefore investigated the potential role of methyl-CpG binding proteins in the regulation of BRCA1 gene expression. In vitro, partial methylation of constructs containing this region strongly inhibits gene expression in the presence of MeCP2 protein. Moreover, in the five human cell lines analyzed, chemically induced hypomethylation is associated with BRCA1 gene activation. These data suggest that methyl-CpG binding proteins might be associated with the control of BRCA1 gene expression and that methyl-DNA binding proteins may participate in the regulation of gene expression in mammalian cells.
Laboratoire de Génétique, UMR 5641 CNRS, UCBL1, and. Laboratoire de Biologie de la Reproduction et du Développement, UCBL1, 69373 Lyon cedex 08, France.
F
F
Magdinier
L M
LM
Billard
G
G
Wittmann
L
L
Frappart
M
M
Benchaïb
G M
GM
Lenoir
J F
JF
Guérin
R
R
Dante
D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D002584:Cervix Uteri / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone; D018899:CpG Islands / Q000235:genetics*; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000077209:Decitabine; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D019398:Genes, BRCA1 / Q000235:genetics*; D005854:Germ Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051783:Methyl-CpG-Binding Protein 2; D011401:Promoter Regions, Genetic / Q000235:genetics; D012097:Repressor Proteins; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection
D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; C497392:MECP2 protein, human; D051783:Methyl-CpG-Binding Protein 2; D012097:Repressor Proteins; D000077209:Decitabine; D001374:Azacitidine