Stage-specific expression of a family of proteins that are major products of zygotic gene activation in the mouse embryo.
Transcriptional activation of the embryonic genome occurs during the two-cell stage in the mouse embryo and is marked by the synthesis of a set of alpha-amanitin-sensitive proteins of Mr 73,000, 70,000, and 68,000. We have characterized these three proteins by two-dimensional gel electrophoresis of [35S]methionine radiolabeled two-cell embryos. Their isoelectric points range from 6.2 to 6.8 and their synthesis, which can constitute 5-10% of total protein synthesis, is restricted to the two-cell stage. These proteins are not heat shock proteins that have previously been reported as major products of transcriptional activation. Peptide mapping by limited proteolysis indicates that these three proteins are highly related to one another and the results of pulse-chase experiments indicate that they are likely to be degraded by the eight-cell stage. These proteins are nuclear-associated and insoluble in 2% Triton X-100/0.3 M KCl. Although these proteins share some features with somatic lamins--they exhibit solubility properties similar to somatic lamins--they do not cross-react with polyclonal antibodies to either lamins A/C or B, nor do they comigrate with somatic lamins on two-dimensional gels. Additional evidence that these proteins are not lamins is that although treatment of two-cell embryos with okadaic acid, which is an inhibitor of protein phosphatases 1 and 2A, results in precocious nuclear envelope breakdown, the proteins remain insoluble in 2% Triton X-100/0.3 M KCl.
Conover
J C
JC
Department of Biology, University of Pennsylvania, Philadelphia 19104-6018.
Temeles
G L
GL
Zimmermann
J W
JW
Burke
B
B
Schultz
R M
RM
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002970:Cleavage Stage, Ovum / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004988:Ethers, Cyclic / Q000494:pharmacology; D015870:Gene Expression; D034882:Lamins; D051379:Mice; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D019319:Okadaic Acid; D010449:Peptide Mapping; D011233:Precipitin Tests; D012995:Solubility
D004988:Ethers, Cyclic; D034882:Lamins; D009687:Nuclear Proteins; D019319:Okadaic Acid