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在杆状病毒系统中合成的沃纳综合征基因产物中的DNA解旋酶活性。

DNA helicase activity in Werner's syndrome gene product synthesized in a baculovirus system.

作者信息

Suzuki N, Shimamoto A, Imamura O, Kuromitsu J, Kitao S, Goto M, Furuichi Y

机构信息

AGENE Research Institute, 200 Kajiwara, Kamakura, Kanagawa 247, Japan.

出版信息

Nucleic Acids Res. 1997 Aug 1;25(15):2973-8. doi: 10.1093/nar/25.15.2973.

Abstract

The gene responsible for Werner's syndrome (WRN) contains a region homologous to the Escherichia coli RecQ type DNA helicase and was thought to code for a DNA helicase belonging to this helicase family. However, no evidence has been shown before to substantiate this prediction. Here, we show data that the product of the WRN gene is indeed a DNA helicase. The gene product, a polypeptide with a relative molecular mass of 170 kDa, expressed in the insect Spodoptera frugiperda (Sf21) cell and purified by affinity column chromatography contained both the ATPase and DNA unwinding activities characteristic of DNA helicase. Expressions in Sf21, as well as in HeLa cells, showed that the WRN DNA helicase is exclusively transported to the nucleoplasm, which is consistent with its function in DNA metabolism. Our studies on strand displacement suggest that WRN helicase can unwind not only a duplex DNA, but also an RNA-DNA heteroduplex, while the latter reaction seems less efficient. Enzymological features learned from the purified WRN helicase are discussed with respect to the biological function, which remains to be clarified.

摘要

导致沃纳综合征(WRN)的基因包含一个与大肠杆菌RecQ型DNA解旋酶同源的区域,曾被认为编码属于该解旋酶家族的一种DNA解旋酶。然而,此前尚无证据证实这一预测。在此,我们展示的数据表明WRN基因的产物确实是一种DNA解旋酶。该基因产物是一种相对分子质量为170 kDa的多肽,在昆虫草地贪夜蛾(Sf21)细胞中表达,并通过亲和柱层析纯化,它兼具DNA解旋酶特有的ATP酶活性和DNA解链活性。在Sf21细胞以及HeLa细胞中的表达表明,WRN DNA解旋酶仅被转运至核质,这与其在DNA代谢中的功能相符。我们对链置换的研究表明,WRN解旋酶不仅可以解开双链DNA,还能解开RNA-DNA异源双链,不过后一种反应似乎效率较低。从纯化的WRN解旋酶中了解到的酶学特性将结合其仍有待阐明的生物学功能进行讨论。

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