Gray M D, Wang L, Youssoufian H, Martin G M, Oshima J
Department of Pathology, University of Washington, Seattle, Washington, 98195-4740, USA.
Exp Cell Res. 1998 Aug 1;242(2):487-94. doi: 10.1006/excr.1998.4124.
Mutations at the Werner helicase locus (WRN) are responsible for the Werner syndrome (WS), a "caricature of aging." We have localized the Werner protein (WRNp) to the nucleoli of replicating mammalian cells, where its appearance is associated with transcriptional activity. A dramatic reduction of the nucleolar signal and of [3H]uridine incorporation occurred when cultures were made quiescent or were exposed to 4-nitroquinoline-1-oxide (4NQO), to which WS cells are particularly susceptible. Total cellular levels of WRNp, however, did not change, and virtually all WRNp was in the nuclear fractions, consistent with translocation to the nucleoplasm and/or masking of the epitopes. The 4NQO-induced altered state of WRNp was prevented by Na3VO4, but not by okadaic acid, suggesting that WRNp localization/function is partially regulated by kinases/phosphatases for Tyr substrates on WRNp or interacting proteins. The repression of rDNA transcription by 4NQO was not reversed by Na3VO4. We suggest that physiological states and genotoxic agents modulate the interaction of WRNp with rDNA, consistent with a role of WRNp in rDNA transcription.
Werner解旋酶基因座(WRN)的突变会导致Werner综合征(WS),这是一种“衰老的典型表现”。我们已将Werner蛋白(WRNp)定位于正在复制的哺乳动物细胞的核仁中,其出现与转录活性相关。当培养物进入静止状态或暴露于WS细胞特别敏感的4-硝基喹啉-1-氧化物(4NQO)时,核仁信号和[3H]尿苷掺入量显著降低。然而,WRNp的总细胞水平没有变化,并且几乎所有WRNp都存在于核组分中,这与向核质的易位和/或表位的掩盖一致。Na3VO4可阻止4NQO诱导的WRNp状态改变,但冈田酸不能,这表明WRNp的定位/功能部分受WRNp或相互作用蛋白上酪氨酸底物的激酶/磷酸酶调节。Na3VO4不能逆转4NQO对rDNA转录的抑制作用。我们认为生理状态和基因毒性剂会调节WRNp与rDNA的相互作用,这与WRNp在rDNA转录中的作用一致。