CDNA CLONING AND SEQUENCING OF HUMAN FIBRILLARIN, A CONSERVED NUCLEOLAR PROTEIN RECOGNIZED BY AUTOIMMUNE ANTISERA
CDNA CLONING AND SEQUENCING OF HUMAN FIBRILLARIN, A CONSERVED NUCLEOLAR PROTEIN RECOGNIZED BY AUTOIMMUNE ANTISERA
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DOI：
10.1073/pnas.88.3.931
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发表时间：
1991-02-01
影响因子：
11.1
通讯作者：
BLOBEL, G
中科院分区：
文献类型：
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作者：
ARIS, JP;BLOBEL, G
我们通过筛选肝癌文库，并与从酿酒酵母（Saccharomyces cerevisiae, NOP1）和非洲爪蟾（Xenopus laevis）的纤维蛋白基因中提取的DNA探针平行，分离出一个编码人纤维蛋白的1.1千碱基cDNA克隆。RNA印迹分析表明，相应的mRNA长度几乎等于1300个核苷酸。体外表达的人纤原蛋白在SDS凝胶上作为一种36kda蛋白迁移，该蛋白被硬皮病自身免疫性疾病患者的抗血清特异性免疫沉淀。人纤维蛋白含有一个长度约为75-80个氨基酸的氨基末端重复结构域，富含甘氨酸和精氨酸残基，与酵母和爪蟾纤维蛋白中的氨基末端结构域相似。在蛋白内出现一个假定的rna结合结构域和一个RNP一致序列，这与纤维蛋白与小核仁rna的关联是一致的。蛋白质序列比对表明，人纤维蛋白中67%的氨基酸与酵母纤维蛋白相同，81%与非洲爪蟾纤维蛋白相同。这种同一性表明，在核糖体生物合成途径的早期，一个重要功能的进化守恒。
We have isolated a 1.1-kilobase cDNA clone that encodes human fibrillarin by screening a hepatoma library in parallel with DNA probes derived from the fibrillarin genes of Saccharomyces cerevisiae (NOP1) and Xenopus laevis. RNA blot analysis indicates that the corresponding mRNA is almost-equal-to 1300 nucleotides in length. Human fibrillarin expressed in vitro migrates on SDS gels as a 36-kDa protein that is specifically immunoprecipitated by antisera from humans with scleroderma autoimmune disease. Human fibrillarin contains an amino-terminal repetitive domain almost-equal-to 75-80 amino acids in length that is rich in glycine and arginine residues and is similar to amino-terminal domains in the yeast and Xenopus fibrillarins. The occurrence of a putative RNA-binding domain and an RNP consensus sequence within the protein is consistent with the association of fibrillarin with small nucleolar RNAs. Protein sequence alignments show that 67% of amino acids from human fibrillarin are identical to those in yeast fibrillarin and that 81% are identical to those in Xenopus fibrillarin. This identity suggests the evolutionary conservation of an important function early in the pathway for ribosome biosynthesis.