Aizawa Youichi, Ishii Susumu
Department of Biology, School of Education, Waseda University, 1-6-1 Nishi-waseda, Shinjuku-ku, 169-8050 Tokyo, Japan.
Gen Comp Endocrinol. 2003 Jul;132(3):465-73. doi: 10.1016/s0016-6480(03)00103-5.
Reptilia is the only vertebrate class in which cDNA for the gonadotropin beta subunit precursor molecule has not been cloned. We have isolated the full-length cDNA clone encoding the LH beta subunit precursor molecule and a partial cDNA clone encoding the FSH beta subunit precursor molecule from a pituitary cDNA library of Reeves's turtle. We further clarified the nucleotide sequence of the remaining part of the turtle FSH beta cDNA and that of full-length cDNA encoding the LH beta subunit precursor molecule of the Japanese grass lizard, by means of the 5' rapid amplification of cDNA end (RACE) and 3' RACE. The nucleotide sequence of the turtle FSH beta cDNA we determined was 584 bp long and contained the coding sequence, 5' untranslated region (UTR) and 3' UTR of 396, 34, and 154 bp, respectively. The nucleotide sequence of the turtle LH beta we isolated was 498 bp long and contained the coding sequence, 5' UTR and 3' UTR of 420, 7, and 71 bp, respectively. The nucleotide sequence of the lizard LH beta we determined was 537 bp long and contained the coding sequence, 5' UTR and 3' UTR of 441, 35, and 61 bp, respectively. Amino acid sequences deduced from coding regions of the turtle FSH beta, LH beta and the lizard LH beta were 131, 139, and 146 residues, respectively. Referring to the amino acid sequences of the bullfrog FSH and LH beta subunit molecules determined chemically, we deduced the amino acid sequences of mature peptide. Amino acid sequences of mature peptides of the turtle FSH, turtle LH, and the lizard LH were 111, 112, and 112 residues, respectively. Amino acid sequences of the mature peptides were compared with those of other vertebrates. The amino acid sequence of the turtle FSH beta subunit molecule was 84.7-85.6, 67.8-71.4, and 61.3-62.2% identical to the FSH sequence of birds, mammals, and amphibians, respectively. The amino acid sequence of the turtle LH beta subunit molecule was 51.6-54.6, 36.2-48.7, and 56.3-57.5% identical to the LH sequence of birds, mammals, and amphibians, respectively. The amino acid sequence of the lizard LH beta subunit molecule was 39.1-47.1, 32.9-43.0, and 46.0-47.3% identical to the LH sequence of birds, mammals, and amphibians, respectively. These identity values suggest that the turtle or reptilian FSH beta subunit molecule is more closely related to avian and mammalian FSH beta subunit molecules than to amphibian FSH beta subunit molecules but reptilian LH beta subunit molecules are more closely related to amphibian LH beta subunit molecules than to avian and mammalian LH beta subunit molecules. This discrepancy in the molecular similarity relationship found in the reptilian FSH and LH beta subunit molecules can be interpreted by assuming that evolution speed was not the same among hormone species and also among vertebrate groups.
爬行纲是唯一尚未克隆出促性腺激素β亚基前体分子cDNA的脊椎动物纲。我们从乌龟的垂体cDNA文库中分离出了编码促黄体生成素(LH)β亚基前体分子的全长cDNA克隆以及编码促卵泡激素(FSH)β亚基前体分子的部分cDNA克隆。我们通过5' cDNA末端快速扩增(RACE)和3' RACE进一步明确了乌龟FSHβ cDNA其余部分的核苷酸序列以及日本草蜥LHβ亚基前体分子全长cDNA的核苷酸序列。我们测定的乌龟FSHβ cDNA的核苷酸序列长584 bp,分别包含396、34和154 bp的编码序列、5'非翻译区(UTR)和3' UTR。我们分离出的乌龟LHβ的核苷酸序列长498 bp,分别包含420、7和71 bp的编码序列、5' UTR和3' UTR。我们测定的蜥蜴LHβ的核苷酸序列长537 bp,分别包含441、35和61 bp的编码序列、5' UTR和3' UTR。从乌龟FSHβ、LHβ以及蜥蜴LHβ的编码区推导的氨基酸序列分别为131、139和146个残基。参照化学测定的牛蛙FSH和LHβ亚基分子的氨基酸序列,我们推导了成熟肽的氨基酸序列。乌龟FSH、乌龟LH以及蜥蜴LH成熟肽的氨基酸序列分别为111、112和112个残基。将成熟肽的氨基酸序列与其他脊椎动物的进行了比较。乌龟FSHβ亚基分子的氨基酸序列与鸟类、哺乳动物和两栖动物FSH序列的同一性分别为84.7 - 85.6%、67.8 - 71.4%和61.3 - 62.2%。乌龟LHβ亚基分子的氨基酸序列与鸟类、哺乳动物和两栖动物LH序列的同一性分别为51.6 - 54.6%、36.2 - 48.7%和56.3 - 57.5%。蜥蜴LHβ亚基分子的氨基酸序列与鸟类、哺乳动物和两栖动物LH序列的同一性分别为39.1 - 47.1%、32.9 - 43.0%和46.0 - 47.3%。这些同一性值表明,乌龟或爬行纲的FSHβ亚基分子与鸟类和哺乳动物FSHβ亚基分子的关系比与两栖动物FSHβ亚基分子的关系更密切,但爬行纲LHβ亚基分子与两栖动物LHβ亚基分子的关系比与鸟类和哺乳动物LHβ亚基分子的关系更密切。在爬行纲FSH和LHβ亚基分子中发现的这种分子相似性关系的差异可以通过假设激素种类之间以及脊椎动物类群之间的进化速度不同来解释。