title: Hydropathy profiles of predicted thyrotropin-releasing hormone precursors are highly conserved despite low similarity of primary structures.
abstract: Two types of cDNAs encoding thyrotropin-releasing hormone (TRH) precursors (TRH-A and TRH-B) were amplified from hypothalamic mRNA of sockeye salmon by reverse transcriptase-polymerase chain reaction (RT-PCR). The amplification was achieved using two primers which correspond to TRH progenitor sequence (Lys/Arg-Arg-Gln-His-Pro-Gly-Lys/Arg-Arg). A full length cDNA encoding TRH-A was obtained by 5'- and 3'-RACE methods. It has a length of 1324 base pairs (bp) that contains sequences of 5' and 3' untranslated regions and an open reading frame of 259 codons. The sockeye salmon TRH-A deduced from the nucleotide sequence tandemly contains 8 copies of TRH progenitor sequences. Another cDNA which encodes a part of TRH-B consists of 242 bp, and the sequence homology between TRH-A and -B cDNAs is 90%. The result of Southern blot analysis of sockeye and masu salmon genomic DNAs supported the evidence that there are at least two TRH genes in the salmonid. A RT-PCR analysis of TRH gene expression in various tissues of sockeye salmon showed that strong expression was observed only in the brain. The primary structure of the sockeye salmon TRH-A shares low similarity to those of human, rat and Xenopus TRH precursors (35, 27 and 44%, respectively). However, their hydropathy profiles were almost the same with each other. The profile of sockeye salmon TRH-A showed the presence of two discrete hydrophobic regions, one in the N-terminal region which corresponds to the signal peptide and the other in the C-terminal region. All of the repetitive TRH progenitor sequences are included in three hydrophilic regions easily recognizable. The present results thus suggest that the three-dimensional structures of TRH precursors are highly conserved, although the primary structures of TRH precursors have diverged through the evolutionary pathway of vertebrates.
authors.affiliation: Department of Applied Biochemistry, Utsunomiya University, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Ohide
authors.forename: H
authors.initials: H
authors.lastname: Ando
authors.forename: T
authors.initials: T
authors.lastname: Yanagisawa
authors.forename: A
authors.initials: A
authors.lastname: Urano
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification*; D019143:Evolution, Molecular; D005260:Female; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000737:chemistry* / Q000235:genetics*; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012474:Salmon; D013973:Thyrotropin-Releasing Hormone / Q000737:chemistry* / Q000235:genetics*; D014158:Transcription, Genetic; D014981:Xenopus
chemical_list: D018076:DNA, Complementary; D011498:Protein Precursors; D012333:RNA, Messenger; D013973:Thyrotropin-Releasing Hormone; C046418:pro-thyrotropin releasing hormone; D011761:Pyrrolidonecarboxylic Acid