A novel sea urchin nuclear receptor encoded by alternatively spliced maternal RNAs.
Screening of a genomic library from the sea urchin Strongylocentrotus purpuratus with a human COUP-TF I cDNA probe revealed the presence of a novel gene member of the steroid-thyroid-retinoic acid receptor superfamily, which was named SpSHR2 (S. purpuratus Steroid Hormone Receptor 2). Sequence analysis of the isolated genomic clone revealed that the DNA binding domain of this orphan receptor is most homologous to the human TR2 receptor. Using this sea urchin genomic fragment as probe, a S. purpuratus embryonic cDNA library was screened and two distinct but homologous cDNA clones were isolated. The two cDNAs encode the same DNA binding domain as the SpSHR2 gene and carry an almost identical 3'-untranslated sequence. One of the clones, however, is missing an entire region of about 1100 nt which includes the putative ligand binding domain. Genomic DNA hybridization suggests that SpSHR2 is a single-copy gene in the S. purpuratus genome. Exon skipping during splicing of a single primary transcript appears to be the reason for the differently sized mRNAs. RNA blot hybridization results suggest that SpSHR2 transcripts are stored as maternal RNA in the egg and are not detected beyond the blastula stage. In vitro transcription and translation of the full-length cDNA produced a polypeptide which specifically binds to the hormone response element in the 5'-flanking region of the sea urchin actin CyIIIb gene. In vivo labeling of proteins synthesized by cleavage stage embryos followed by immune precipitation of the SpSHR2 protein using specific antibodies reveals that the maternal SpSHR2 mRNA is being translated during early embryonic development.
Kontrogianni-Konstantopoulos
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Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
Vlahou
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Vu
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Flytzanis
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D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D051838:COUP Transcription Factor I; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis*; D004625:Embryo, Nonmammalian / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D015698:Genomic Library; D008297:Male; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics*; D011965:Receptors, Glucocorticoid / Q000235:genetics*; D012617:Sea Urchins / Q000196:embryology / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000096:biosynthesis*; D014158:Transcription, Genetic
D051838:COUP Transcription Factor I; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; D014157:Transcription Factors