Recognition of the CDEI motif GTCACATG by mouse nuclear proteins and interference with the early development of the mouse embryo.
We have reported previously (1) two unexpected consequences of the microinjection into fertilized mouse eggs of a recombinant plasmid designated p12B1, carrying a 343 bp insert of non-repetitive mouse DNA. Injected at very low concentrations, this plasmid could be established as an extrachromosomal genetic element. When injected in greater concentration, an early arrest of embryonic development resulted. In the present work, we have studied this toxic effect in more detail by microinjecting short synthetic oligonucleotides with sequences from the mouse insert. Lethality was associated with the nucleotide sequence GTCACATG, identical with the CDEl element of yeast centromeres. Development of injected embryos was arrested between the one-cell and the early morula stages, with abnormal structures and DNA contents. Electrophoretic mobility shift and DNAse foot-printing assays demonstrated the binding of mouse nuclear protein(s) to the CDEl-like box. Base changes within the CDEl sequence prevented both the toxic effects in embryos and the formation of protein complex in vitro, suggesting that protein binding at such sites in chromosomal DNA plays an important role in early development.
Blangy
A
A
Unité 273 de I'INSERM, Université de Nice-Sophia Antipolis, France.
Léopold
P
P
Vidal
F
F
Rassoulzadegan
M
M
Cuzin
F
F
D000818:Animals; D001483:Base Sequence; D002845:Chromatography; D005314:Embryonic and Fetal Development; D051379:Mice; D008845:Microinjections; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009687:Nuclear Proteins / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D010957:Plasmids / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*
D009687:Nuclear Proteins; D009841:Oligonucleotides