3'-Half of the thrombopoietin cDNA confers higher expression of erythropoietin at the RNA level but not at the protein level.
Both erythropoietin (EPO) and the short-form thrombopoietin (TPO) were expressed at low levels whereas the long-form TPO was expressed at high levels in transgenic animals. To elucidate the role of carboxy-terminal half of the long-form TPO which is absent in the short-form, we generated recombinant TPO or EPO expression vectors which contain or lack the carboxy-terminal half of TPO and examined their expression in the HC11 and 293 cells. The long-form TPO was expressed higher than the short-form regardless of the cell types, transfection modes, and promoters. When 3'-half of the long-form TPO cDNA was placed downstream of the EPO cDNA to act as a 3'-untranslated region, expression of EPO was moderately increased at the RNA level, however, no remarkable increase was observed at the protein level. These results suggest that the low expression of EPO, as like as the short-form TPO, is due to absence of the 3'-half in the full-length TPO that confers stability both at the RNA and protein levels.
Kim
Tae-Won
TW
Department of Biology, Dongguk University, Seoul 100-715, Korea.
Ji
Jin Woo
JW
Chang
Ho Gun
HG
Kim
Myoung Ok
MO
Ryoo
Zae Young
ZY
Park
In Kook
IK
Kim
Sun Jung
SJ
D020413:3' Untranslated Regions / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002364:Caseins / Q000235:genetics; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics; D018076:DNA, Complementary / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D004921:Erythropoietin / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics; D012313:RNA / Q000378:metabolism*; D020871:RNA Stability / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013926:Thrombopoietin / Q000235:genetics*
D020413:3' Untranslated Regions; D002364:Caseins; D018076:DNA, Complementary; D011994:Recombinant Proteins; D004921:Erythropoietin; D012313:RNA; D013926:Thrombopoietin