Transcriptional repression of p27 is essential for murine embryonic development.

Youichi Teratake,Masayasu Kitahashi,Lisa Fujimura,Haruko Watanabe-Takano,Akemi Sakamoto,Yoshiharu Sato,Yuta Hasegawa,Masahiko Hatano,Chisa Kuga,Masafumi Arima,Takeshi Tokuhisa

doi:10.1038/srep26244

Youichi Teratake, Masayasu Kitahashi + Show 9 more

Open Access

https://doi.org/10.1038/srep26244

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Journal: Scientific Reports	Publication Date: May 1, 2016
Citations: 10	License type: CC BY 4.0

Affiliation: Chiba University, Chiba University Hospital

Abstract

The Nczf gene has been identified as one of Ncx target genes and encodes a novel KRAB zinc-finger protein, which functions as a sequence specific transcriptional repressor. In order to elucidate Nczf functions, we generated Nczf knockout (Nczf−/−) mice. Nczf−/− mice died around embryonic day 8.5 (E8.5) with small body size and impairment of axial rotation. Histopathological analysis revealed that the cell number decreased and pyknotic cells were occasionally observed. We examined the expression of cell cycle related genes in Nczf−/− mice. p27 expression was increased in E8.0 Nczf−/− mice compared to that of wild type mice. Nczf knockdown by siRNA resulted in increased expression of p27 in mouse embryonic fibroblasts (MEFs). Furthermore, p27 promoter luciferase reporter gene analysis confirmed the regulation of p27 mRNA expression by Nczf. Nczf−/−; p27−/− double knockout mice survived until E11.5 and the defect of axial rotation was restored. These data suggest that p27 repression by Nczf is essential in the developing embryo.

Highlights

Cell proliferation and differentiation are coordinated during development
To determine the stage at which Nczf knockout (Nczf−/−)embryos died during embryonic development, embryos were isolated from timed heterozygous intercrosses from embryonic day 14.5 (E14.5) to as early as E6.5
Since p27 mRNA expression is elevated in E8.5 Nczf−/−embryos and the elevation may cause the impairment of cell proliferation and embryonic lethality, we generated Nczf−/−; p27−/−double knockout mice to examine the effect of p27 in the Nczf−/−genotype

Summary

Introduction

Cell proliferation and differentiation are coordinated during development. Chromosome status is monitored at the G1 and G2 cell cycle checkpoints to start DNA replication and cell division, respectively. Histological analysis revealed that Nczf−/−embryos at E6.5 and E7.5 were smaller than wild type mice (Fig. 2B). To investigate the molecular link between Nczf deficiency and cell proliferation, we examined the expression of cell cycle regulator genes in Nczf−/−embryos.

Results

Conclusion