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  • 标题:Charged residues in the H-NS linker drive DNA binding and gene silencing in single cells
  • 本地全文:下载
  • 作者:Yunfeng Gao ; Yong Hwee Foo ; Ricksen S. Winardhi
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:2017
  • 卷号:114
  • 期号:47
  • 页码:12560-12565
  • DOI:10.1073/pnas.1716721114
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:Nucleoid-associated proteins (NAPs) facilitate chromosome organization in bacteria, but the precise mechanism remains elusive. H-NS is a NAP that also plays a major role in silencing pathogen genes. We used genetics, single-particle tracking in live cells, superresolution microscopy, atomic force microscopy, and molecular dynamics simulations to examine H-NS/DNA interactions in single cells. We discovered a role for the unstructured linker region connecting the N-terminal oligomerization and C-terminal DNA binding domains. In the present work we demonstrate that linker amino acids promote engagement with DNA. In the absence of linker contacts, H-NS binding is significantly reduced, although no change in chromosome compaction is observed. H-NS is not localized to two distinct foci; rather, it is scattered all around the nucleoid. The linker makes DNA contacts that are required for gene silencing, while chromosome compaction does not appear to be an important H-NS function.
  • 关键词:H-NS ; nucleoid-associated proteins ; superresolution microscopy ; single-particle tracking ; atomic force microscopy
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