期刊
NUCLEIC ACIDS RESEARCH
卷 38, 期 9, 页码 3019-3030出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkp1221
关键词
-
资金
- Ministry of Education, Culture, Sports, Science, and Technology (MEXT)
- Japan Society for the Promotion of Science (JSPS) [19657002, 21370001, 19.07420]
- EMBO
- National Institutes of Health [1R01GM081680-01]
- 21st century COE project
- Polish Academy of Sciences
- University of Tokyo
- Grants-in-Aid for Scientific Research [19657002, 21370001] Funding Source: KAKEN
- Direct For Biological Sciences
- Div Of Biological Infrastructure [0832858] Funding Source: National Science Foundation
Genome comparison and genome context analysis were used to find a putative mobile element in the genome of Photorhabdus luminescens, an entomopathogenic bacterium. The element is composed of 16-bp direct repeats in the terminal regions, which are identical to a part of insertion sequences (ISs), a DNA methyltransferase gene homolog, two genes of unknown functions and an open reading frame (ORF) (plu0599) encoding a protein with no detectable sequence similarity to any known protein. The ORF (plu0599) product showed DNA endonuclease activity, when expressed in a cell-free expression system. Subsequently, the protein, named R.PluTI, was expressed in vivo, purified and found to be a novel type IIF restriction enzyme that recognizes 5'-GGCGC/C-3' (/ indicates position of cleavage). R.PluTI cleaves a two-site supercoiled substrate at both the sites faster than a one-site supercoiled substrate. The modification enzyme homolog encoded by plu0600, named M.PluTI, was expressed in Escherichia coli and shown to protect DNA from R.PluTI cleavage in vitro, and to suppress the lethal effects of R.PluTI expression in vivo. These results suggested that they constitute a restriction-modification system, present on the putative mobile element. Our approach thus allowed detection of a previously uncharacterized family of DNA-interacting proteins.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据