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整合宿主因子在IS50和Tn5转座过程中发挥作用。

Integration host factor plays a role in IS50 and Tn5 transposition.

作者信息

Makris J C, Nordmann P L, Reznikoff W S

机构信息

Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin-Madison 53706.

出版信息

J Bacteriol. 1990 Mar;172(3):1368-73. doi: 10.1128/jb.172.3.1368-1373.1990.

Abstract

In Escherichia coli, the frequencies of IS50 and Tn5 transposition are greater in Dam- cells than in isogenic Dam+ cells. IS50 transposition is increased approximately 1,000-fold and Tn5 transposition frequencies are increased about 5- to 10-fold in the absence of Dam methylation. However, in cells that are deficient for both integration host factor (IHF) and Dam methylase, the transposition frequencies of IS50 and Tn5 approximate those found in wild-type cells. The absence of IHF alone has no effect on either IS50 or Tn5 transposition. These results suggest that IHF is required for the increased transposition frequencies of IS50 and Tn5 that are observed in Dam- cells. It is also shown that the level of expression of IS50-encoded proteins, P1 and P2, required for IS50 and Tn5 transposition and its regulation does not decrease in IHF- or in IHF- Dam- cells. This result suggests that the effects of IHF on IS50 and Tn5 transposition are not at the level of IS50 gene expression. Finally, IHF is demonstrated to significantly retard the electrophoretic mobility of a 289-base-pair segment of IS50 DNA that contains a putative IHF protein-binding site. The physiological role of this IHF binding site remains to be determined.

摘要

在大肠杆菌中,IS50和Tn5的转座频率在Dam-细胞中比在同基因的Dam+细胞中更高。在缺乏Dam甲基化的情况下,IS50转座增加约1000倍,Tn5转座频率增加约5至10倍。然而,在整合宿主因子(IHF)和Dam甲基酶均缺陷的细胞中,IS50和Tn5的转座频率接近野生型细胞中的频率。单独缺乏IHF对IS50或Tn5转座均无影响。这些结果表明,在Dam-细胞中观察到的IS50和Tn5转座频率增加需要IHF。还表明,IS50和Tn5转座及其调控所需的IS50编码蛋白P1和P2的表达水平在IHF-或IHF-Dam-细胞中并未降低。这一结果表明,IHF对IS50和Tn5转座的影响不在IS50基因表达水平。最后,证明IHF能显著减缓包含假定IHF蛋白结合位点的IS50 DNA的289个碱基对片段的电泳迁移率。该IHF结合位点的生理作用仍有待确定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1188/208607/b6f14e522c00/jbacter01045-0217-a.jpg

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