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端粒酶具有持续性。

Telomerase is processive.

作者信息

Greider C W

机构信息

Cold Spring Harbor Laboratory, New York 11724.

出版信息

Mol Cell Biol. 1991 Sep;11(9):4572-80. doi: 10.1128/mcb.11.9.4572-4580.1991.

Abstract

Telomerase synthesizes tandem repeats of the sequence d(TTGGGG) onto input d(TTGGGG)n primer oligonucleotides (C. W. Greider and E. H. Blackburn, Cell 43:405-413). An intrinsic RNA component of the enzyme provides the template for d(TTGGGG)n repeat synthesis [C. W. Greider and E. H. Blackburn, Nature (London) 337:331-337, 1989; G.-L. Lu, J. D. Bradley, L. D. Attardi, and E. H. Blackburn, Nature (London) 344:126-132, 1990]. In a typical reaction, products greater than 2,000 nucleotides were synthesized in 60 min. Dilution and primer challenge experiments showed that these long products were synthesized processively. The apparent processivity was not due to a higher affinity of the enzyme for long d(TTGGGG) products over the shorter competitors. The degree of processivity was quantitated; telomerase synthesized approximately 520 nucleotides before half of the enzyme had dissociated. After dissociating, telomerase reinitiated d(TTGGGG)n synthesis on new primer oligonucleotides. The products from a telomerase reaction have a characteristic 6-nucleotide banding pattern (C. W. Greider and E. H. Blackburn, Cell 51:887-898, 1987). A strong pause in the reaction occurs after the addition of the first G in the sequence d(TTGGGG). Both the processivity and the banding pattern analysis imply that in the elongation mechanism there must be a translocation step after the 9 nucleotides of internal template RNA have been copied to the extreme 5' end.

摘要

端粒酶将序列d(TTGGGG)的串联重复序列合成到输入的d(TTGGGG)n引物寡核苷酸上(C.W.格雷德和E.H.布莱克本,《细胞》43:405 - 413)。该酶的一个内在RNA组分提供了d(TTGGGG)n重复序列合成的模板[C.W.格雷德和E.H.布莱克本,《自然》(伦敦)337:331 - 337,1989;G.-L.卢、J.D.布拉德利、L.D.阿塔尔迪和E.H.布莱克本,《自然》(伦敦)344:126 - 132,1990]。在典型反应中,60分钟内合成了大于2000个核苷酸的产物。稀释和引物挑战实验表明,这些长产物是连续合成的。明显的持续性并非由于该酶对长d(TTGGGG)产物的亲和力高于较短的竞争产物。持续性程度进行了定量;端粒酶在一半的酶解离之前合成了大约520个核苷酸。解离后,端粒酶在新的引物寡核苷酸上重新启动d(TTGGGG)n的合成。端粒酶反应的产物具有特征性的6核苷酸条带模式(C.W.格雷德和E.H.布莱克本,《细胞》51:887 - 898,1987)。在序列d(TTGGGG)中添加第一个G后,反应会出现强烈的停顿。持续性和条带模式分析都表明,在延伸机制中,内部模板RNA的9个核苷酸被复制到最末端的5'端之后,必定存在一个易位步骤。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/352a/361337/265cc17b2596/molcellb00033-0298-a.jpg

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