The zebrafish erythropoietin: functional identification and biochemical characterization.
In the present study, the zebrafish epo cDNA was cloned. The encoded protein displays 90%, 55% and 32% identity to the Epo from carp, fugu and human, respectively. Through RT-PCR, 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 alphaA1- and betaA1-globin, but not the embryonic betae1-globin, was affected.
Chu
Cheng-Ying
CY
Graduate Institute of Biochemical Sciences, National Taiwan University, Taipei 106, Taiwan.
Cheng
Chia-Hsiung
CH
Chen
Gen-Der
GD
Chen
Yi-Chung
YC
Hung
Chin-Chun
CC
Huang
Kai-Yun
KY
Huang
Chang-Jen
CJ
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D041905:Erythroid Cells / Q000378:metabolism; D004921:Erythropoietin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D006454:Hemoglobins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009928:Organ Specificity; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D006454:Hemoglobins; D012333:RNA, Messenger; D029961:Zebrafish Proteins; C523494:epoa protein, zebrafish; D004921:Erythropoietin