Localization and differential expression of the Krüppel-associated box zinc finger proteins 1 and 54 in early mouse development.
Upon fertilization, the zygotic genome is activated. To ensure the transcription of specific genes and avoid promiscuous gene expression, a chromatin-mediated repressive state is established. To characterize potential heterochromatin factors present during the first cleavage, two putative transcriptional repressors, zinc finger protein (ZFP1) and ZFP54, belonging to the Kr&#x00FC;ppel-associated box (KRAB) zinc finger family, were isolated. ZFP1 and ZFP54 contain an N-terminally located KRAB repressor domain followed by 8 and 12 repeats of Kr&#x00FC;ppel zinc-finger motifs, respectively. Reverse transcription (RT) and quantitative (q) PCR show that maternally contributed Zfp1 and Zfp54 mRNA are detected throughout preimplantation development. &#x03B1;-Amanitin-treated zygotes revealed that maternal Zfp1 and Zfp54 are fully degraded at the two-cell stage. Microinjections of in vitro-transcribed mRNA encoding a gfp-fused reporter gene into zygotes demonstrated the intracellular distribution of ZFP1-green fluorescent protein (GFP) and ZFP54-GFP colocalized with a DNA marker in the two-cell embryo. The KRAB domain was essential to colocalize with DNA, and deletion of the KRAB domain in ZFP1-GFP and ZFP54-GFP localized in nucleoli and in a ubiquitously manner, respectively. Taken together, this suggests a role for ZFP1 and ZFP54 in transcriptional regulation in early development.
Albertsen
Maria
M
Department of Medical Biochemistry, Aarhus University, Aarhus C, Denmark.
Teperek
Marta
M
Elholm
Grethe
G
Füchtbauer
Ernst-Martin
EM
Lykke-Hartmann
Karin
K
D053959:Alpha-Amanitin / Q000494:pharmacology; D000818:Animals; D001755:Blastocyst; D002467:Cell Nucleus / Q000235:genetics*; D003433:Crosses, Genetic; D004622:Embryo, Mammalian; D005786:Gene Expression Regulation; D051741:Kruppel-Like Transcription Factors; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics* / Q000502:physiology
D053959:Alpha-Amanitin; D051741:Kruppel-Like Transcription Factors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C553210:zinc finger protein 1, mouse; C553211:zinc finger protein 54, mouse