The zebrafish erythropoietin: Functional identification and biochemical characterization

被引:52
作者
Chu, Cheng-Ying
Cheng, Chia-Hsiung
Chen, Gen-Der
Chen, Yi-Chung
Hung, Chin-Chun
Huang, Kai-Yun
Huang, Chang-Jen
机构
[1] Acad Sinica, Inst Biol Chem, Taipei 115, Taiwan
[2] Natl Taiwan Univ, Grad Inst Biochem Sci, Taipei 106, Taiwan
[3] Acad Sinica, Isnt Cellular & Organism Biol, Taipei 115, Taiwan
关键词
erythropoietin; zebrafish; morpholino oligonucleotide; glycoprotein; secretion; globin staining;
D O I
10.1016/j.febslet.2007.07.073
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In the present study, the zebratish epo cDNA was cloned. The encoded protein displays 90%, 55% and 32% identity to the Epo from carp, fugu and human, respectively. Through RTPCR, the expression of zepo mRNA was mainly in the heart and liver. In the COS-1 cell transfection experiments, the recombinant zEpo-HA protein was efficiently secreted into the culture medium as a glycoprotein and the carbohydrate moiety can be cleaved by the treatment of peptide-N-glycosidase F (PNGase F). Using the morpholino approach, we showed that zepo morphants displayed severe anemia leading to high mortality during development. Such an effect can be significantly rescued by zepo RNA. Furthermore, in the absence of functional zEpo, the expression of specific markers for adult globin genes, such as alpha A1- and beta A1-globin, but not the embryonic beta e1-globin, was affected. (c) 2007 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:4265 / 4271
页数:7
相关论文
共 22 条
[1]  
BOISSEL JP, 1993, J BIOL CHEM, V268, P15983
[2]   EORTC guidelines for the use of erythropoietic proteins in anaemic patients with cancer [J].
Bokemeyer, C ;
Aapro, MS ;
Courdi, A ;
Foubert, J ;
Link, H ;
Österborg, A ;
Repetto, L ;
Soubeyran, P .
EUROPEAN JOURNAL OF CANCER, 2004, 40 (15) :2201-2216
[3]   Characterization of adult alpha- and beta-globin genes in the zebrafish [J].
Chan, FY ;
Robinson, J ;
Brownlie, A ;
Shivdasani, RA ;
Donovan, A ;
Brugnara, C ;
Kim, J ;
Lau, BC ;
Witkowska, HE ;
Zon, LI .
BLOOD, 1997, 89 (02) :688-700
[4]   Requirement of nuclear localization and transcriptional activity of p53 for its targeting to the yolk syncytial layer (YSL) nuclei in zebrafish embryo and its use for apoptosis assay [J].
Chen, GD ;
Chou, CM ;
Hwang, SPL ;
Wang, FF ;
Chen, YC ;
Hung, CC ;
Chen, JY ;
Huang, CJ .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2006, 344 (01) :272-282
[5]   Erythropoietin gene from a teleost fish, Fugu rubripes [J].
Chou, CF ;
Tohari, S ;
Brenner, S ;
Venkatesh, B .
BLOOD, 2004, 104 (05) :1498-1503
[6]   ROLE OF GLYCOSYLATION ON THE SECRETION AND BIOLOGICAL-ACTIVITY OF ERYTHROPOIETIN [J].
DELORME, E ;
LORENZINI, T ;
GIFFIN, J ;
MARTIN, F ;
JACOBSEN, F ;
BOONE, T ;
ELLIOTT, S .
BIOCHEMISTRY, 1992, 31 (41) :9871-9876
[7]   INTRAEMBRYONIC HEMATOPOIETIC-CELL MIGRATION DURING VERTEBRATE DEVELOPMENT [J].
DETRICH, HW ;
KIERAN, MW ;
CHAN, FY ;
BARONE, LM ;
YEE, K ;
RUNDSTADLER, JA ;
PRATT, S ;
RANSOM, D ;
ZON, LI .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (23) :10713-10717
[8]   STRUCTURAL REQUIREMENTS FOR ADDITION OF O-LINKED CARBOHYDRATE TO RECOMBINANT ERYTHROPOIETIN [J].
ELLIOTT, S ;
BARTLEY, T ;
DELORME, E ;
DERBY, P ;
HUNT, R ;
LORENZINI, T ;
PARKER, V ;
ROHDE, MF ;
STONEY, K .
BIOCHEMISTRY, 1994, 33 (37) :11237-11245
[9]   Beneficial and ominous aspects of the pleiotropic action of erythropoietin [J].
Jelkmann, W ;
Wagner, K .
ANNALS OF HEMATOLOGY, 2004, 83 (11) :673-686
[10]   ERYTHROPOIETIN - STRUCTURE, CONTROL OF PRODUCTION, AND FUNCTION [J].
JELKMANN, W .
PHYSIOLOGICAL REVIEWS, 1992, 72 (02) :449-489