Identification, Characterization, and Effects of Xenopus laevis PNAS-4 Gene on Embryonic Development

Abstract EN

Apoptosis plays an important role in embryonic development.

PNAS-4 has been demonstrated to induce apoptosis in several cancer cells.

In this study, we cloned Xenopus laevis PNAS-4 (xPNAS-4), which is homologous to the human PNAS-4 gene.

Bioinformatics analysis for PNAS-4 indicated that xPNAS-4 shared 87.6% identity with human PNAS-4 and 85.5% with mouse PNAS-4.

The phylogenetic tree of PNAS-4 protein was also summarized.

An analysis of cellular localization using an EGFP-fused protein demonstrated that xPNAS-4 was localized in the perinuclear region of the cytoplasm.

RT-PCR analysis revealed that xPNAS-4, as a maternally expressed gene, was present in all stages of early embryo development.

Whole-mount in situ hybridization showed that xPNAS-4 was mainly expressed in ectoderm and mesoderm.

Furthermore, microinjection of xPNAS-4 mRNA in vivo caused developmental defects manifesting as a small eye phenotype in the Xenopous embryos, and as a small eye or one-eye phenotype in developing zebrafish embryos.

In addition, embryos microinjected with xPNAS-4 antisense morpholino oligonucleotides (MOs) exhibited a failure of head development and shortened axis.