Characterization of a novel cDNA obtained through differential-display PCR of phorbol ester-stimulated ovarian tissue from the brook trout (Salvelinus fontinalis).
A cDNA (DRC1, differentially regulated clone 1) was obtained from differential-display polymerase chain reaction (PCR) of brook trout ovarian tissue stimulated with phorbol-12-myristate-13-acetate (PMA) and A23187. Using 5' RACE (rapid amplification of cDNA ends), two full-length clones were obtained from DRC1 that were 425 and 660 base pairs long and contained the same open reading frame. On Northern blots, DRC1 hybridized with two ovarian mRNAs of 0.45 and 0.7 kb that were significantly suppressed in the presence of PMA and/or A23187. The mRNAs were not observed in ovaries prior to the resumption of meiosis but were present during ovulation and 24 hr after ovulation. Of other trout tissues tested by Northern blotting, the expression of DRC1-related transcripts also was extremely high in the liver. Based on the full-length cDNAs obtained from RACE, these mRNAs presumably encode an 88-amino-acid protein (DRTP1, differentially regulated trout protein 1) that is homologous to a gene superfamily composed of snake venom neurotoxins, a CD59 complement regulatory protein, Ly-6 alloantigens, and a urokinase-type plasminogen activator receptor. To our knowledge, this is the first description of this type of cDNA from a nonmammalian source other than snake venom. In view of the sequence homology and tissue expression of DRTP1, a possible function of this protein may be to regulate the complement system in trout.
Lee
P H
PH
Department of Biological Sciences, University of Notre Dame, IN 46556, USA.
Goetz
F W
FW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry* / Q000302:isolation & purification; D005260:Female; D029941:Fish Proteins; D008969:Molecular Sequence Data; D010053:Ovary / Q000187:drug effects* / Q000378:metabolism*; D010703:Phorbol Esters / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification; D012313:RNA / Q000032:analysis; D017386:Sequence Homology, Amino Acid; D014337:Trout / Q000235:genetics*
D018076:DNA, Complementary; D029941:Fish Proteins; D010703:Phorbol Esters; D011506:Proteins; C110691:differentially regulated trout protein 1, Salvelinus fontinalis; D012313:RNA