Identification, characterization, and effects of Xenopus laevis PNAS-4 gene on embryonic development.
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.
Yan
Fei
F
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China.
Ruan
Xu-zhi
XZ
Yang
Han-shuo
HS
Yao
Shao-hua
SH
Zhao
Xin-yu
XY
Gou
Lan-tu
LT
Ma
Fan-xin
FX
Yuan
Zhu
Z
Deng
Hong-xin
HX
Wei
Yu-quan
YQ
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D019295:Computational Biology; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000196:embryology* / Q000378:metabolism; D047108:Embryonic Development / Q000235:genetics*; D005124:Eye Abnormalities / Q000473:pathology; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D008845:Microinjections; D008969:Molecular Sequence Data; D010802:Phylogeny; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D029867:Xenopus Proteins / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics*
D051017:Apoptosis Regulatory Proteins; C524210:DESI2 protein, Xenopus; D012333:RNA, Messenger; D029867:Xenopus Proteins
Methods Cell Biol. 1991;36:685-95
1811161
Cell Death Differ. 1998 Feb;5(2):156-62
10200460
J Biol Chem. 2000 Apr 7;275(14):10484-91
10744739
Cell. 1982 Oct;30(3):675-86
6183003
FEBS Lett. 2008 Jun 25;582(15):2325-32
18538138
Int J Dev Biol. 2000 Aug;44(5):507-10
11032186
Mech Dev. 2003 Mar;120(3):315-23
12591601
Dev Biol. 1999 Sep 1;213(1):142-56
10452852
Genes Dev. 1994 Jun 1;8(11):1285-99
7926731
Oncology. 2008;75(3-4):137-44
18824876
Cell. 1997 Feb 7;88(3):347-54
9039261
Nature. 1992 Mar 19;356(6366):215-21
1552940
Cell Mol Life Sci. 1999 Jan;55(1):38-47
10065150
Cell. 1995 Oct 20;83(2):279-87
7585945
Development. 1990 Nov;110(3):955-62
2088731
Ophthalmic Res. 2004 Jan-Feb;36(1):4-24
15007235
Biochem J. 2001 Nov 15;360(Pt 1):39-47
11695990
J Anat. 2002 Aug;201(2):121-35
12220121
Science. 1995 Mar 10;267(5203):1456-62
7878464
Cell Biol Int. 2009 Mar;33(3):276-82
19101642
Mech Dev. 1997 Dec;69(1-2):183-95
9486540
Cell Tissue Res. 2000 Jul;301(1):43-52
10928280
Biochimie. 2009 Apr;91(4):502-7
19133312
Am J Respir Cell Mol Biol. 1998 Jan;18(1):21-31
9448042
J Am Soc Nephrol. 1994 Jul;5(1):12-21
7948779
Ment Retard Dev Disabil Res Rev. 2001;7(4):261-6
11754520
J Biol Chem. 2005 Jun 10;280(23):22135-45
15826955