Cloning and expression of rat preproendothelin-3 cDNA.
We report here the cloning and expression of a rat full-length cDNA encoding preproendothelin-3 (preproET-3). The predicted rat preproET-3 consisted of 167 amino acid residues. As in other ET-family peptides, the mature rat ET-3 was predicted to be produced through unusual processing from a 41-residue intermediate, the big ET-3 in rat. Transient transfection of COS-7 cells with the cloned preproET-3 cDNA resulted in the production of mature ET-3 and this production was inhibited by phosphoramidon, a metaloprotease inhibitor. This suggested that a phosphoramidon sensitive mechanism was involved in the production of ET-3 in the transfected COS-7 cells. Northern blot analysis showed that an approximately 3.0-kb rat preproET-3 mRNA was expressed in rat tissues, including the eye ball, submandibular gland, brain, kidney, jejunum, stomach and spleen. A 2.0-kb and a 3.3-kb mRNA were also detected in the eye ball and small intestine, respectively. The distinct distribution of rat preproET-3 mRNA from that of preproET-1 mRNA suggested that ET-1 and ET-3 played different roles.
Shiba
R
R
Department of Pharmacology, University of Tsukuba, Ibaraki, Japan.
Sakurai
T
T
Yamada
G
G
Morimoto
H
H
Saito
A
A
Masaki
T
T
Goto
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D016232:Endothelins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D004792:Enzyme Precursors; D015723:Gene Library; D006801:Humans; D007124:Immunoenzyme Techniques; D007421:Intestine, Small / Q000502:physiology; D008969:Molecular Sequence Data; D009799:Ocular Physiological Phenomena; D009838:Oligodeoxyribonucleotides; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D016232:Endothelins; D004792:Enzyme Precursors; D009838:Oligodeoxyribonucleotides; D011498:Protein Precursors; D012333:RNA, Messenger; D011994:Recombinant Proteins; C075625:preproendothelin-3; D011061:Poly A; D012313:RNA; D004247:DNA