Conservation of the egg envelope digestion mechanism of hatching enzyme in euteleostean fishes.
We purified two hatching enzymes, namely high choriolytic enzyme (HCE; EC 3.4.24.67) and low choriolytic enzyme (LCE; EC 3.4.24.66), from the hatching liquid of Fundulus heteroclitus, which were named Fundulus HCE (FHCE) and Fundulus LCE (FLCE). FHCE swelled the inner layer of egg envelope, and FLCE completely digested the FHCE-swollen envelope. In addition, we cloned three Fundulus cDNAs orthologous to cDNAs for the medaka precursors of egg envelope subunit proteins (i.e. choriogenins H, H minor and L) from the female liver. Cleavage sites of FHCE and FLCE on egg envelope subunit proteins were determined by comparing the N-terminal amino acid sequences of digests with the sequences deduced from the cDNAs for egg envelope subunit proteins. FHCE and FLCE cleaved different sites of the subunit proteins. FHCE efficiently cleaved the Pro-X-Y repeat regions into tripeptides to dodecapeptides to swell the envelope, whereas FLCE cleaved the inside of the zona pellucida domain, the core structure of egg envelope subunit protein, to completely digest the FHCE-swollen envelope. A comparison showed that the positions of hatching enzyme cleavage sites on egg envelope subunit proteins were strictly conserved between Fundulus and medaka. Finally, we extended such a comparison to three other euteleosts (i.e. three-spined stickleback, spotted halibut and rainbow trout) and found that the egg envelope digestion mechanism was well conserved among them. During evolution, the egg envelope digestion by HCE and LCE orthologs was established in the lineage of euteleosts, and the mechanism is suggested to be conserved.
Atmosphere and Ocean Research Institute, The University of Tokyo, Chiba, Japan.
Mari
M
Kawaguchi
Shigeki
S
Yasumasu
Akio
A
Shimizu
Kaori
K
Sano
Ichiro
I
Iuchi
Mutsumi
M
Nishida
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D004527:Egg Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005260:Female; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005399:Fishes / Q000235:genetics* / Q000378:metabolism*; D005432:Flounder / Q000235:genetics; D023781:Fundulidae / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000235:genetics; D009990:Oryzias / Q000235:genetics; D010063:Ovum / Q000378:metabolism; D010802:Phylogeny; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D023701:Smegmamorpha / Q000235:genetics
D017931:DNA Primers; D004527:Egg Proteins; D029941:Fish Proteins; C092024:L-SF protein, Oryzias latipes; D011498:Protein Precursors; C104950:choriogenin H; D008666:Metalloendopeptidases; C024344:envelysin