CHARACTERIZATION OF A HIGHLY CONSERVED 23 KDA BASIC PROTEIN

高度保守的 23 KDA 碱性蛋白的表征

基本信息

项目摘要

During the isolation and characterization of cDNA clones for ADP-ribosylation factors, about 20 kDa GTP-binding proteins, a human brain cDNA containing a partial open reading frame encoding 48 amino acids, of which 10 were basic, was isolated. This peptide was unusual based on its predicted pI of 11.7. Oligonucleotide probes specific for this sequence hybridized with two abundant mRNAs in HL-60 cells, one of which appeared to comigrate with an ARF3 mRNA. Because of the unusual nature of this peptide as well as the size and abundance of the mRNAs, studies were performed to identify and investigate the relationship between these mRNAs. The cDNA fragment was used to isolate overlapping clones from dibutyryl cAMP-differentiated HL-60 and bovine retinal cDNA libraries. A putative open reading frame was identified that encodes a 23.6 kDa protein (p23, 203 amino acids) with a predicted pI of 11.6. The deduced amino acid sequence is highly conserved between human and bovine, exhibiting 97% identity; they are 99% identical if conservative substitutions are included. The human and bovine nucleotide sequences exhibit 91% identity across the coding region. Clones from both species contain a poly-adenylation signal (AATAAA) 4 nucleotides downstream (3') from the termination codon as well as a poly(A)+ tail. Other clones, however, extended beyond the point of poly(A)+ addition. In one bovine clone, the 3'-untranslated region extended 442 nucleotides beyond the polyadenylation signal to end in a poly(A)+ tail; a second polyadenylation signal was found 21 nucleotides before the poly (A)+ tail. Hybridization of poly(A)+ RNA from various bovine tissues and brains of several mammalian species with cDNA and oligonucleotides specific for the coding region identified two abundant mRNA species of 1.2- and 0.8-kb. Oligonucleotide probes specific for the extended 3'-untranslated region hybridized with the 1.2-kb, but not the 0.8-kb, mRNA. Both mRNAs were observed in all mammalian tissues and species. These studies indicate that a ubiquitous, basic 23.6 kDa protein is highly conserved among mammals and is encoded by two mRNA species that appear to differ in the site of polyadenylation.
在分离和表征用于 ADP-核糖基化因子,约20 kDa GTP结合蛋白,人脑 包含编码48个氨基酸的部分开放阅读框的cDNA 其中10个是碱性的。这种肽是不寻常的, 预测pI为11.7。特异于该序列的寡核苷酸探针 与HL-60细胞中两种丰富的mRNA杂交,其中一种似乎 与ARF 3 mRNA共迁移。由于这种肽的特殊性质 以及mRNA的大小和丰度,进行了研究, 鉴定并研究这些mRNA之间的关系。的cDNA 片段用于从二丁酰 cAMP分化的HL-60和牛视网膜cDNA文库。推定的 鉴定了一个开放阅读框,其编码23.6 kDa蛋白(p23,203 氨基酸),预测pI为11.6。推导的氨基酸序列 在人和牛之间高度保守,表现出97%的同一性;它们 如果包括保守替换,则99%相同。的人力和 牛核苷酸序列在整个编码区表现出91%的同一性。 来自两个物种的克隆都含有多聚腺苷酸化信号(AATAAA)4 终止密码子下游(3 ')的核苷酸以及 poly(A)+ tail。然而,其他克隆延伸到poly(A)+点之外 的增订条文在一个牛克隆中,3 '-非翻译区延伸442 多聚腺苷酸化信号之外的核苷酸以聚腺苷酸+尾结束; 在poly(A)+前21个核苷酸处发现第二个多聚腺苷酸化信号 尾巴不同牛组织和脑中poly(A)+ RNA的杂交 几种哺乳动物的cDNA和寡核苷酸特异性, 编码区鉴定了1.2和0.8 kb的两种丰富的mRNA种类。 特异于延伸的3 '-非翻译区的寡核苷酸探针 与1.2kb的mRNA杂交,而不与0.8kb的mRNA杂交。两种mRNA均为 在所有哺乳动物组织和物种中观察到。这些研究表明 一种普遍存在的碱性23.6kDa蛋白在哺乳动物中高度保守, 由两种mRNA编码,这两种mRNA似乎在 聚腺苷酸化

项目成果

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S R PRICE其他文献

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{{ truncateString('S R PRICE', 18)}}的其他基金

GENES FOR GTP-BINDING PROTEINS
GTP 结合蛋白的基因
  • 批准号:
    3919999
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:
CHARACTERIZATION OF NATIVE AND RECOMBINANT ADP-RIBOSYLATION FACTORS
天然和重组 ADP-核糖化因子的表征
  • 批准号:
    3857988
  • 财政年份:
  • 资助金额:
    --
  • 项目类别:

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胰岛素1、2缺失小鼠骨骼肌克隆细胞的建立及功能分析
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