Molecular cloning of a cDNA that encodes the precursor to several exogastrula-inducing peptides, epidermal-growth-factor-related polypeptides of the sea urchin Anthocidaris crassispina.
Complementary DNA clones for exogastrula-inducing peptides (EGIPs) of the sea urchin Anthocidaris crassispina, which are related to epidermal growth factor (EGF), were obtained from a cDNA library of late gastrula embryos using, as probe, the partial cDNA for one of the EGIP (EGIP-D) obtained by the reverse-transcription PCR method. The longest cDNA was composed of 1662 bp, and encoded a protein of approximately 36 kDa with a region that resembled a signal sequence. The deduced protein contains the sequences of EGIP-C, EGIP-D, and EGIP-A in that order, followed by the sequence for an unidentified EGIP-like polypeptide. When expressed in Escherichia coli as a fusion protein with beta-galactosidase, the product for the cDNA was specifically recognized by a rabbit antibody raised against EGIP-D that had been purified from embryos. Characteristic amino acid residues were found around the N-terminus and the C-terminus of each EGIP sequence, suggesting a specific processing mechanism for the generation of the individual EGIPs from the precursor. RNA-blot analysis revealed the presence of EGIP mRNA in unfertilized eggs. The level of this mRNA decreased gradually after fertilization, began to increase dramatically after the onset of gastrulation, and continued to increase through the pluteus stage. Genomic Southern-blot analysis suggested that this gene is present as a single copy. A homology search showed that the EGIP cDNA has a similarity to the cDNA for SpEGF2 which was cloned as a gastrula-specific gene in another sea urchin, Strongylocentrotus purpuratus.
Yamasu
K
K
Department of Regulation Biology, Faculty of Science, Saitama University, Urawa, Japan.
Watanabe
H
H
Kohchi
C
C
Soma
G
G
Mizuno
D
D
Akasaka
K
K
Shimada
H
H
Suyemitsu
T
T
Ishihara
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D000013:Congenital Abnormalities / Q000209:etiology*; D018076:DNA, Complementary / Q000737:chemistry; D004815:Epidermal Growth Factor / Q000235:genetics* / Q000502:physiology; D005775:Gastrula / Q000502:physiology*; D007447:Invertebrate Hormones / Q000235:genetics*; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D012617:Sea Urchins / Q000196:embryology*
D018076:DNA, Complementary; D007447:Invertebrate Hormones; D011498:Protein Precursors; D012333:RNA, Messenger; C061708:exogastrula-inducing peptide A, Anthocidaris crassispina; C061709:exogastrula-inducing peptide D, Anthocidaris crassispina; C061707:exogastrula-inducing peptide C, Anthocidaris crassispina; D004815:Epidermal Growth Factor