Structure of the RTP-DNA complex and the mechanism of polar replication fork arrest

J. A. Wilce; J. P. Vivian; A. F. Hastings; G. Otting; R. H.A. Folmer; I. G. Duggin; R. G. Wake; M. C.J. Wilce

doi:10.1038/84934

Structure of the RTP-DNA complex and the mechanism of polar replication fork arrest

J. A. Wilce, J. P. Vivian, A. F. Hastings, G. Otting, R. H.A. Folmer, I. G. Duggin, R. G. Wake, M. C.J. Wilce^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

31 Citations (Scopus)

Abstract

The coordinated termination of DNA replication is an important step in the life cycle of bacteria with circular chromosomes, but has only been defined at a molecular level in two systems to date. Here we report the structure of an engineered replication terminator protein (RTP) ot Bacillus subtilis in complex with a 21 base pair DNA by X-ray crystallography at 2.5 Å resolution. We also use NMR spectroscopic titration techniques. This work reveals a novel DNA interaction involving a dimeric 'winged helix' domain protein that differs from predictions. While the two recognition helices of RTP ate in close contact with the B-form DNA major grooves, the 'wings' and N-termini of RTP do not form intimate contacts with the DNA. This structure provides insight into the molecular basis of polar replication fork arrest based on a model of cooperative binding and differential binding affinities of RTP to the two adjacent binding sites in file complete terminator.

Original language	English
Pages (from-to)	206-210
Number of pages	5
Journal	Nature Structural Biology
Volume	8
Issue number	3
DOIs	https://doi.org/10.1038/84934
Publication status	Published - 2001
Externally published	Yes

Access to Document

10.1038/84934

Cite this

@article{059206709b9642bdb97201e3f40571cc,

title = "Structure of the RTP-DNA complex and the mechanism of polar replication fork arrest",

abstract = "The coordinated termination of DNA replication is an important step in the life cycle of bacteria with circular chromosomes, but has only been defined at a molecular level in two systems to date. Here we report the structure of an engineered replication terminator protein (RTP) ot Bacillus subtilis in complex with a 21 base pair DNA by X-ray crystallography at 2.5 {\AA} resolution. We also use NMR spectroscopic titration techniques. This work reveals a novel DNA interaction involving a dimeric 'winged helix' domain protein that differs from predictions. While the two recognition helices of RTP ate in close contact with the B-form DNA major grooves, the 'wings' and N-termini of RTP do not form intimate contacts with the DNA. This structure provides insight into the molecular basis of polar replication fork arrest based on a model of cooperative binding and differential binding affinities of RTP to the two adjacent binding sites in file complete terminator.",

author = "Wilce, \{J. A.\} and Vivian, \{J. P.\} and Hastings, \{A. F.\} and G. Otting and Folmer, \{R. H.A.\} and Duggin, \{I. G.\} and Wake, \{R. G.\} and Wilce, \{M. C.J.\}",

year = "2001",

doi = "10.1038/84934",

language = "English",

volume = "8",

pages = "206--210",

journal = "Nature Structural Biology",

issn = "1072-8368",

publisher = "Nature Research",

number = "3",

}

TY - JOUR

T1 - Structure of the RTP-DNA complex and the mechanism of polar replication fork arrest

AU - Wilce, J. A.

AU - Vivian, J. P.

AU - Hastings, A. F.

AU - Otting, G.

AU - Folmer, R. H.A.

AU - Duggin, I. G.

AU - Wake, R. G.

AU - Wilce, M. C.J.

PY - 2001

Y1 - 2001

N2 - The coordinated termination of DNA replication is an important step in the life cycle of bacteria with circular chromosomes, but has only been defined at a molecular level in two systems to date. Here we report the structure of an engineered replication terminator protein (RTP) ot Bacillus subtilis in complex with a 21 base pair DNA by X-ray crystallography at 2.5 Å resolution. We also use NMR spectroscopic titration techniques. This work reveals a novel DNA interaction involving a dimeric 'winged helix' domain protein that differs from predictions. While the two recognition helices of RTP ate in close contact with the B-form DNA major grooves, the 'wings' and N-termini of RTP do not form intimate contacts with the DNA. This structure provides insight into the molecular basis of polar replication fork arrest based on a model of cooperative binding and differential binding affinities of RTP to the two adjacent binding sites in file complete terminator.

AB - The coordinated termination of DNA replication is an important step in the life cycle of bacteria with circular chromosomes, but has only been defined at a molecular level in two systems to date. Here we report the structure of an engineered replication terminator protein (RTP) ot Bacillus subtilis in complex with a 21 base pair DNA by X-ray crystallography at 2.5 Å resolution. We also use NMR spectroscopic titration techniques. This work reveals a novel DNA interaction involving a dimeric 'winged helix' domain protein that differs from predictions. While the two recognition helices of RTP ate in close contact with the B-form DNA major grooves, the 'wings' and N-termini of RTP do not form intimate contacts with the DNA. This structure provides insight into the molecular basis of polar replication fork arrest based on a model of cooperative binding and differential binding affinities of RTP to the two adjacent binding sites in file complete terminator.

UR - https://www.scopus.com/pages/publications/0035122946

U2 - 10.1038/84934

DO - 10.1038/84934

M3 - Article

SN - 1072-8368

VL - 8

SP - 206

EP - 210

JO - Nature Structural Biology

JF - Nature Structural Biology

IS - 3

ER -

Structure of the RTP-DNA complex and the mechanism of polar replication fork arrest

Abstract

Access to Document

Other files and links

Fingerprint

Cite this