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蛋白质HU在大肠杆菌染色体复制起点的酶促复制过程中的作用

Protein HU in the enzymatic replication of the chromosomal origin of Escherichia coli.

作者信息

Dixon N E, Kornberg A

出版信息

Proc Natl Acad Sci U S A. 1984 Jan;81(2):424-8. doi: 10.1073/pnas.81.2.424.

Abstract

A protein that stimulates the enzymatic replication of duplex DNAs of recombinant phages and plasmids bearing the Escherichia coli origin of replication (oriC) has been isolated from an extract of E. coli. The isolated protein and the well-known protein HU, a histone-like DNA-binding protein, have identical polypeptide molecular weights and saturate the oriC replication assay at less than 40 dimers per template DNA circle. This level is one-tenth that needed to coat the template. Protein HU from the blue-green alga Anabaena is similarly active. Antibody specific for protein HU from E. coli inhibits replication promoted both by the reconstituted system and by a crude enzyme extract; in both assays, activity is restored by excess of the isolated protein. Cells lysed in 1 M KCl yield 32,000 dimers of the protein per cell, a number consistent with the reported abundance of HU. These data establish the identity of the isolated factor and protein HU and provide an indication of a function for HU in replication.

摘要

一种能刺激带有大肠杆菌复制起点(oriC)的重组噬菌体和质粒双链DNA进行酶促复制的蛋白质已从大肠杆菌提取物中分离出来。分离出的这种蛋白质与著名的蛋白质HU(一种类组蛋白DNA结合蛋白)具有相同的多肽分子量,并且在每个模板DNA环少于40个二聚体时就能使oriC复制检测达到饱和。这个水平是覆盖模板所需水平的十分之一。来自蓝藻鱼腥藻的蛋白质HU也有类似活性。针对大肠杆菌蛋白质HU的特异性抗体能抑制重组系统和粗酶提取物所促进的复制;在这两种检测中,过量的分离蛋白可恢复活性。在1M KCl中裂解的细胞每个细胞产生32,000个该蛋白质的二聚体,这个数量与报道的HU丰度一致。这些数据确定了分离因子与蛋白质HU的同一性,并为HU在复制中的功能提供了一个线索。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7649/344689/3fc03be87a92/pnas00603-0145-a.jpg

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