Newgard C B, Nakano K, Hwang P K, Fletterick R J
Proc Natl Acad Sci U S A. 1986 Nov;83(21):8132-6. doi: 10.1073/pnas.83.21.8132.
We have cloned the cDNA encoding glycogen phosphorylase (1,4-alpha-D-glucan:orthophosphate alpha-D-glucosyl-transferase, EC 2.4.1.1) from human liver. Blot-hybridization analysis using a large fragment of the cDNA to probe mRNA from rabbit brain, muscle, and liver tissues shows preferential hybridization to liver RNA. Determination of the entire nucleotide sequence of the liver message has allowed a comparison with the previously determined rabbit muscle phosphorylase sequence. Despite an amino acid identity of 80%, the two cDNAs exhibit a remarkable divergence in G+C content. In the muscle phosphorylase sequence, 86% of the nucleotides at the third codon position are either deoxyguanosine or deoxycytidine residues, while in the liver homolog the figure is only 60%, resulting in a strikingly different pattern of codon usage throughout most of the sequence. The liver phosphorylase cDNA appears to represent an evolutionary mosaic; the segment encoding the N-terminal 80 amino acids contains greater than 90% G+C at the third codon position. A survey of other published mammalian cDNA sequences reveals that the data for liver and muscle phosphorylases reflects a bias in codon usage patterns in liver and muscle coding sequences in general.
我们已从人肝脏中克隆出编码糖原磷酸化酶(1,4-α-D-葡聚糖:正磷酸α-D-葡糖基转移酶,EC 2.4.1.1)的cDNA。使用该cDNA的大片段对兔脑、肌肉和肝脏组织的mRNA进行印迹杂交分析,结果显示与肝脏RNA有优先杂交。对肝脏信使RNA的完整核苷酸序列进行测定后,得以与先前测定的兔肌肉磷酸化酶序列进行比较。尽管氨基酸同一性为80%,但这两个cDNA在G+C含量上表现出显著差异。在肌肉磷酸化酶序列中,第三个密码子位置86%的核苷酸为脱氧鸟苷或脱氧胞苷残基,而在肝脏同源物中这一比例仅为60%,导致在大部分序列中密码子使用模式明显不同。肝脏磷酸化酶cDNA似乎代表了一种进化镶嵌体;编码N端80个氨基酸的片段在第三个密码子位置的G+C含量大于90%。对其他已发表的哺乳动物cDNA序列进行的一项调查显示,肝脏和肌肉磷酸化酶的数据总体上反映了肝脏和肌肉编码序列中密码子使用模式的偏差。