[LagoZ and LagZ, 2 genes depleted of CpG dinucleotides, derived from the LacZ gene for the study of epigenetic control].
The methylation of 5'CpG 3' dinucleotides within genes creates potential targets for protein complexes that bind to methylated DNA sequences and to histone deacetylases (MBD-HDAC). This can lead to transcriptional repression by modification of chromatic. To test the importance of this repression in vivo and to determine when during development these epigenetic controls are placed on genes, two novel genes have been engineered by directed mutagenesis of the CpG-rich LacZ gene that are depleted of (LagZ) or completely lacking (LagoZ) CpG sequences. We report that the expression (transcriptional and translational) of the three genes is indistinguishable in transient assays in cleaving mouse embryos. Therefore, the complete absence of CpG sequences within three kilobases of coding sequence is compatible with its maintenance in the nucleus and with its expression. These molecules can now be used to study the ontogenesis of the CpG-dependent repressive system in intact organisms.
Henry
I
I
Unité de biologie moléculaire du développement, Institut Pasteur, Paris, France.
Forlani
S
S
Vaillant
S
S
Muschler
J
J
Choulika
A
A
Nicolas
J F
JF
D000107:Acetylation; D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000378:metabolism; D018899:CpG Islands; D019175:DNA Methylation; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005813:Genes, Synthetic; D006657:Histones / Q000378:metabolism; D007763:Lac Operon; D051379:Mice; D008845:Microinjections; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic / Q000235:genetics; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012097:Repressor Proteins / Q000502:physiology; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis
D006657:Histones; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D001616:beta-Galactosidase