Document Detail

Expression and genomic organization of a medaka fish novel membrane form of guanylyl cyclase/orphan receptor.
MedLine Citation:
PMID:  12777830     Owner:  NLM     Status:  MEDLINE    
A novel membrane guanylyl cyclase (membrane GC), OlGC8, was identified in the medaka fish Oryzias latipes by the isolation of full-length cDNA (4958 bp) and genomic DNA (14.3 kbp) clones. Phylogenetic analysis indicated that OlGC8 does not belong in any known vertebrate membrane GC subfamily. OlGC8 consists of an extracellular domain (214 residues), a transmembrane segment (19 residues), and an intracellular protein kinase-like domain (284 residues) and a cyclase catalytic domain (228 residues), although the extracellular domain is about half the length (around 450 residues) of other known vertebrate membrane GCs. OlGC8 transiently expressed in COS-7 cells exhibited only basal guanylyl cyclase activity. None of the known ligands (rat ANP, BNP, CNP, and C-ANF) and various medaka fish tissue extracts, which activated OlGC1, OlGC2, and OlGC7 differentially, stimulated basal activity, suggesting that OlGC8 is an orphan receptor. The OlGC8 gene consists of 24 exons and exists as a single copy on the medaka fish genome. Northern blot hybridization showed that a 5 kb-OlGC8 mRNA was expressed in the kidney and the testis at a high level and a 3.3 kb-OlGC8 mRNA was expressed only in the brain. The RNase protection, RNA Ligase-Mediated Rapid Amplification of cDNA Ends (RLM-RACE), and reverse transcription-polymerase chain reaction (RT-PCR) analyses demonstrated that the 3.3 kb-OlGC8 mRNA detected in the brain is transcribed from the second transcription initiation site, and contains an intron at the position prior to the catalytic domain, the translation product of which appears to be a protein lacking the cyclase catalytic domain.
Sayaka Yamagami; Shan Hua Xu; Makiko Tsutsumi; Hiroshi Hori; Norio Suzuki
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Zoological science     Volume:  20     ISSN:  0289-0003     ISO Abbreviation:  Zool. Sci.     Publication Date:  2003 May 
Date Detail:
Created Date:  2003-06-02     Completed Date:  2003-09-29     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8702287     Medline TA:  Zoolog Sci     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  591-606     Citation Subset:  IM    
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.
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MeSH Terms
Amino Acid Sequence
Base Sequence
Blotting, Southern
COS Cells
Cloning, Molecular
DNA Primers
DNA, Complementary / analysis
Gene Expression Profiling
Guanylate Cyclase / chemistry,  genetics*,  metabolism
Membrane Proteins / chemistry,  genetics*,  metabolism
Molecular Sequence Data
Oryzias / genetics*
Receptors, Atrial Natriuretic Factor / chemistry,  genetics*,  metabolism
Reverse Transcriptase Polymerase Chain Reaction
Sequence Alignment
Reg. No./Substance:
0/DNA Primers; 0/DNA, Complementary; 0/Membrane Proteins; EC Cyclase; EC, Atrial Natriuretic Factor

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