Abstract
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.
Original language | English (US) |
---|---|
Pages (from-to) | 12560-12565 |
Number of pages | 6 |
Journal | Proceedings of the National Academy of Sciences of the United States of America |
Volume | 114 |
Issue number | 47 |
DOIs | |
State | Published - Nov 21 2017 |
Externally published | Yes |
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Keywords
- Atomic force microscopy
- H-NS
- Nucleoid-associated proteins
- Single-particle tracking
- Superresolution microscopy
ASJC Scopus subject areas
- General
Cite this
Charged residues in the H-NS linker drive DNA binding and gene silencing in single cells. / Gao, Yunfeng; Foo, Yong Hwee; Winardhi, Ricksen S.; Tang, Qingnan; Yan, Jie; Kenney, Linda.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, No. 47, 21.11.2017, p. 12560-12565.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Charged residues in the H-NS linker drive DNA binding and gene silencing in single cells
AU - Gao, Yunfeng
AU - Foo, Yong Hwee
AU - Winardhi, Ricksen S.
AU - Tang, Qingnan
AU - Yan, Jie
AU - Kenney, Linda
PY - 2017/11/21
Y1 - 2017/11/21
N2 - 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.
AB - 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.
KW - Atomic force microscopy
KW - H-NS
KW - Nucleoid-associated proteins
KW - Single-particle tracking
KW - Superresolution microscopy
UR - http://www.scopus.com/inward/record.url?scp=85034585453&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85034585453&partnerID=8YFLogxK
U2 - 10.1073/pnas.1716721114
DO - 10.1073/pnas.1716721114
M3 - Article
C2 - 29109287
AN - SCOPUS:85034585453
VL - 114
SP - 12560
EP - 12565
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 47
ER -