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异质核糖核蛋白A2/B1亚型与端粒单链DNA的相互作用及体外功能

Interaction of hnRNP A2/B1 isoforms with telomeric ssDNA and the in vitro function.

作者信息

Kamma H, Fujimoto M, Fujiwara M, Matsui M, Horiguchi H, Hamasaki M, Satoh H

机构信息

Institute of Basic Medical Sciences, University of Tsukuba, Ibaraki 305-8575, Japan.

出版信息

Biochem Biophys Res Commun. 2001 Jan 26;280(3):625-30. doi: 10.1006/bbrc.2000.4173.

Abstract

Overexpression of heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1, especially of B1 has been reported as a useful marker to detect cancers in early stage, although the biological reason is not clear. A2/B1 proteins were previously reported to bind telomeric DNA repeats. Alternative splicing of A2/B1 gene produces abundant A2, less abundant B1, and testis-specific minor isoforms B0a and B0b. In this study, B1 and B0b that have the N-terminal 12 amino acid insertion were suggested to have higher affinities to telomeric single-stranded DNA (ssDNA) than A2 and B0a. Kinetic analyses using purified B1 and B0b indicated that they interact dynamically with a single array of telomeric repeats. Furthermore, functional assays demonstrated that B1 and B0b bind with telomeric repeats in a tandem fashion and protect them from a nuclease and promote telomerase activity. A2/B1 proteins, especially B1 and B0b, may function as telomeric ssDNA-binding proteins in cancer and reproductive cells.

摘要

据报道,异质性核糖核蛋白(hnRNP)A2/B1的过表达,尤其是B1的过表达,是检测早期癌症的一个有用标志物,尽管其生物学原因尚不清楚。此前有报道称,A2/B1蛋白可结合端粒DNA重复序列。A2/B1基因的可变剪接产生大量的A2、少量的B1以及睾丸特异性的次要异构体B0a和B0b。在本研究中,具有N端12个氨基酸插入的B1和B0b对端粒单链DNA(ssDNA)的亲和力被认为高于A2和B0a。使用纯化的B1和B0b进行的动力学分析表明,它们与端粒重复序列的单阵列动态相互作用。此外,功能分析表明,B1和B0b以串联方式与端粒重复序列结合,保护它们免受核酸酶的作用,并促进端粒酶活性。A2/B1蛋白,尤其是B1和B0b,可能在癌症和生殖细胞中作为端粒ssDNA结合蛋白发挥作用。

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