Krieger M, Coge F, Gros F, Thibault J
Laboratoire de Biochimie Cellulaire, Collège de France, Paris.
Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2161-5. doi: 10.1073/pnas.88.6.2161.
A cDNA clone for dopa decarboxylase (EC 4.1.1.28) has been isolated from a rat pheochromocytoma cDNA library and the cDNA sequence has been determined. It corresponds to an mRNA of 2094 nucleotides. The length of the mRNA was measured by primer-extension of rat pheochromocytoma RNA and the 5' end of the sequence of the mRNA was confirmed by the PCR. A probe spanning the translation initiation site of the mRNA was used to hybridize with mRNAs from various organs of the rat. S1 nuclease digestion of the mRNAs annealed with this probe revealed two classes of mRNAs. The comparison of the cDNA sequence and published sequences for rat liver, human pheochromocytoma, and Drosophila dopa decarboxylase supported the conclusion that two mRNAs are produced: one is specific for tissue of neuronal origin and the other is specific for tissues of nonneuronal (mesodermal or endodermal) origin. The neuronal mRNA contains a 5' untranslated sequence that is highly conserved between human and rat pheochromocytoma including a GA stretch. The coding sequence and the 3' untranslated sequence of mRNAs from rat liver and pheochromocytoma are identical. The rat mRNA differs only in the 5' untranslated region. Thus a unique gene codes for dopa decarboxylase and this gene gives rise to at least two transcripts presumably in response to different signals during development.
已从大鼠嗜铬细胞瘤cDNA文库中分离出多巴脱羧酶(EC 4.1.1.28)的cDNA克隆,并测定了该cDNA序列。它对应于一个2094个核苷酸的mRNA。通过对大鼠嗜铬细胞瘤RNA进行引物延伸来测量mRNA的长度,并通过PCR确认mRNA序列的5'端。使用跨越mRNA翻译起始位点的探针与大鼠各器官的mRNA杂交。用该探针退火的mRNA经S1核酸酶消化后显示出两类mRNA。将该cDNA序列与已发表的大鼠肝脏、人嗜铬细胞瘤和果蝇多巴脱羧酶的序列进行比较,支持了这样的结论:产生了两种mRNA:一种对神经源性组织具有特异性,另一种对非神经源性(中胚层或内胚层)组织具有特异性。神经mRNA包含一个5'非翻译序列,该序列在人和大鼠嗜铬细胞瘤之间高度保守,包括一段GA序列。大鼠肝脏和嗜铬细胞瘤的mRNA的编码序列和3'非翻译序列是相同的。大鼠mRNA仅在5'非翻译区域有所不同。因此,一个独特的基因编码多巴脱羧酶,并且该基因可能在发育过程中响应不同信号而产生至少两种转录本。