TcpA from the Clostridium perfringens plasmid pCW3 is more closely related to the DNA translocase FtsK than to coupling proteins

Daouda A.K. Traore; Von Vergel L. Torres; Naureen Akhtar; Alexandra M. Gummer; Sarena F. Flanigan; Fasséli Coulibaly; Vicki Adams; James C. Whisstock; Julian I. Rood

doi:10.1016/j.str.2023.02.001

TcpA from the Clostridium perfringens plasmid pCW3 is more closely related to the DNA translocase FtsK than to coupling proteins

Daouda A.K. Traore, Von Vergel L. Torres, Naureen Akhtar, Alexandra M. Gummer, Sarena F. Flanigan, Fasséli Coulibaly, Vicki Adams, James C. Whisstock (Leading Author), Julian I. Rood (Leading Author)

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

Abstract

Conjugative DNA transfer is a major factor in the dissemination of antibiotic resistance and virulence genes. In the Gram-positive pathogen Clostridium perfringens, the majority of conjugative plasmids share the conserved tcp locus that governs the assembly of the transfer system. Here, we describe multiple structures of the coupling protein TcpA, an essential ATPase that is suggested to provide the mechanical force to propel the DNA through the transfer apparatus. The structures of TcpA in the presence and absence of nucleotides revealed conformational rearrangements and highlight a crucial role for the unstructured C terminus. Our findings reveal that TcpA shares most structural similarity with the FtsK DNA translocase, a central component of the bacterial cell division machinery. Our structural data suggest that conjugation in C. perfringens may have evolved from the bacterial chromosome segregation system and, accordingly, suggest the possibility that double-stranded DNA is transferred through the Tcp conjugation apparatus.

Original language	English
Pages (from-to)	455-463.e4
Number of pages	14
Journal	Structure
Volume	31
Issue number	4
DOIs	https://doi.org/10.1016/j.str.2023.02.001
Publication status	Published - Apr 2023

Keywords

ATPase
Clostridium perfringens
conjugation
coupling protein
FtsK
pCW3
TcpA
translocase

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10.1016/j.str.2023.02.001Licence: CC BY

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@article{0ea2922f5354405eb6304bd129f1c56c,

title = "TcpA from the Clostridium perfringens plasmid pCW3 is more closely related to the DNA translocase FtsK than to coupling proteins",

abstract = "Conjugative DNA transfer is a major factor in the dissemination of antibiotic resistance and virulence genes. In the Gram-positive pathogen Clostridium perfringens, the majority of conjugative plasmids share the conserved tcp locus that governs the assembly of the transfer system. Here, we describe multiple structures of the coupling protein TcpA, an essential ATPase that is suggested to provide the mechanical force to propel the DNA through the transfer apparatus. The structures of TcpA in the presence and absence of nucleotides revealed conformational rearrangements and highlight a crucial role for the unstructured C terminus. Our findings reveal that TcpA shares most structural similarity with the FtsK DNA translocase, a central component of the bacterial cell division machinery. Our structural data suggest that conjugation in C. perfringens may have evolved from the bacterial chromosome segregation system and, accordingly, suggest the possibility that double-stranded DNA is transferred through the Tcp conjugation apparatus.",

keywords = "ATPase, Clostridium perfringens, conjugation, coupling protein, FtsK, pCW3, TcpA, translocase",

author = "Traore, {Daouda A.K.} and Torres, {Von Vergel L.} and Naureen Akhtar and Gummer, {Alexandra M.} and Flanigan, {Sarena F.} and Fass{\'e}li Coulibaly and Vicki Adams and Whisstock, {James C.} and Rood, {Julian I.}",

note = "Funding Information: This research was supported by Project Grant GNT1104502 from the Australian National Health and Medical Research Council awarded to J.I.R. D.A.K.T. and J.C.W. We thank the CSIRO Collaborative Crystallization Center (www.csiro.au/C3), Melbourne, Australia. We thank the Monash Macromolecular Crystallisation Facility (MMCF) and the Monash Proteomic Plat-form Clayton, Australia. X-ray diffraction experiments were undertaken on the MX beamlines at the Australian Synchrotron, part of ANSTO, and we thank, in particular, Tom Caradoc-Davies and Santosh Panjikar, staff members of the MX beamlines, for their assistance with data collection and processing. Conceptualization, D.A.K.T. V.V.L.T. N.A. F.C. V.A. J.C.W. and J.I.R.; investigation, D.A.K.T. V.V.L.T. N.A. A.M.G. S.F.F. and F.C.; methodology, D.A.K.T. V.V.L.T. N.A. A.M.G. S.F.F. and F.C.; formal analysis, D.A.K.T. V.V.L.T. N.A. A.M.G. S.F.F. F.C. V.A. J.C.W. and J.I.R.; funding acquisition, D.A.K.T. J.C.W. and J.I.R.; project administration, D.A.K.T. V.A. J.C.W. and J.I.R.; resources, D.A.K.T. V.A. J.C.W. and J.I.R.; validation, D.A.K.T. F.C. and V.A.; data curation, D.A.K.T. and F.C.; supervision, D.A.K.T. V.A. J.C.W. and J.I.R.; visualization, D.A.K.T. and F.C.; writing – original draft, D.A.K.T. V.A. J.C.W. and J.I.R.; writing – review & editing, D.A.K.T. V.V.L.T. N.A. A.M.G. S.F.F. F.C. V.A. J.C.W. and J.I.R. The authors declare no competing interests. We support inclusive, diverse, and equitable conduct of research. One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in their field of research or within their geographical location. One or more of the authors of this paper self-identifies as living with a disability. Funding Information: This research was supported by Project Grant GNT1104502 from the Australian National Health and Medical Research Council awarded to J.I.R., D.A.K.T., and J.C.W. We thank the CSIRO Collaborative Crystallization Center ( www.csiro.au/C3 ), Melbourne, Australia. We thank the Monash Macromolecular Crystallisation Facility (MMCF) and the Monash Proteomic Plat-form Clayton, Australia. X-ray diffraction experiments were undertaken on the MX beamlines at the Australian Synchrotron, part of ANSTO, and we thank, in particular, Tom Caradoc-Davies and Santosh Panjikar, staff members of the MX beamlines, for their assistance with data collection and processing. Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2023",

month = apr,

doi = "10.1016/j.str.2023.02.001",

language = "English",

volume = "31",

pages = "455--463.e4",

journal = "Structure",

issn = "0969-2126",

publisher = "Cell Press",

number = "4",

}

TY - JOUR

T1 - TcpA from the Clostridium perfringens plasmid pCW3 is more closely related to the DNA translocase FtsK than to coupling proteins

AU - Traore, Daouda A.K.

AU - Torres, Von Vergel L.

AU - Akhtar, Naureen

AU - Gummer, Alexandra M.

AU - Flanigan, Sarena F.

AU - Coulibaly, Fasséli

AU - Adams, Vicki

A2 - Whisstock, James C.

A2 - Rood, Julian I.

N1 - Funding Information: This research was supported by Project Grant GNT1104502 from the Australian National Health and Medical Research Council awarded to J.I.R. D.A.K.T. and J.C.W. We thank the CSIRO Collaborative Crystallization Center (www.csiro.au/C3), Melbourne, Australia. We thank the Monash Macromolecular Crystallisation Facility (MMCF) and the Monash Proteomic Plat-form Clayton, Australia. X-ray diffraction experiments were undertaken on the MX beamlines at the Australian Synchrotron, part of ANSTO, and we thank, in particular, Tom Caradoc-Davies and Santosh Panjikar, staff members of the MX beamlines, for their assistance with data collection and processing. Conceptualization, D.A.K.T. V.V.L.T. N.A. F.C. V.A. J.C.W. and J.I.R.; investigation, D.A.K.T. V.V.L.T. N.A. A.M.G. S.F.F. and F.C.; methodology, D.A.K.T. V.V.L.T. N.A. A.M.G. S.F.F. and F.C.; formal analysis, D.A.K.T. V.V.L.T. N.A. A.M.G. S.F.F. F.C. V.A. J.C.W. and J.I.R.; funding acquisition, D.A.K.T. J.C.W. and J.I.R.; project administration, D.A.K.T. V.A. J.C.W. and J.I.R.; resources, D.A.K.T. V.A. J.C.W. and J.I.R.; validation, D.A.K.T. F.C. and V.A.; data curation, D.A.K.T. and F.C.; supervision, D.A.K.T. V.A. J.C.W. and J.I.R.; visualization, D.A.K.T. and F.C.; writing – original draft, D.A.K.T. V.A. J.C.W. and J.I.R.; writing – review & editing, D.A.K.T. V.V.L.T. N.A. A.M.G. S.F.F. F.C. V.A. J.C.W. and J.I.R. The authors declare no competing interests. We support inclusive, diverse, and equitable conduct of research. One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in their field of research or within their geographical location. One or more of the authors of this paper self-identifies as living with a disability. Funding Information: This research was supported by Project Grant GNT1104502 from the Australian National Health and Medical Research Council awarded to J.I.R., D.A.K.T., and J.C.W. We thank the CSIRO Collaborative Crystallization Center ( www.csiro.au/C3 ), Melbourne, Australia. We thank the Monash Macromolecular Crystallisation Facility (MMCF) and the Monash Proteomic Plat-form Clayton, Australia. X-ray diffraction experiments were undertaken on the MX beamlines at the Australian Synchrotron, part of ANSTO, and we thank, in particular, Tom Caradoc-Davies and Santosh Panjikar, staff members of the MX beamlines, for their assistance with data collection and processing. Publisher Copyright: © 2023 The Author(s)

PY - 2023/4

Y1 - 2023/4

N2 - Conjugative DNA transfer is a major factor in the dissemination of antibiotic resistance and virulence genes. In the Gram-positive pathogen Clostridium perfringens, the majority of conjugative plasmids share the conserved tcp locus that governs the assembly of the transfer system. Here, we describe multiple structures of the coupling protein TcpA, an essential ATPase that is suggested to provide the mechanical force to propel the DNA through the transfer apparatus. The structures of TcpA in the presence and absence of nucleotides revealed conformational rearrangements and highlight a crucial role for the unstructured C terminus. Our findings reveal that TcpA shares most structural similarity with the FtsK DNA translocase, a central component of the bacterial cell division machinery. Our structural data suggest that conjugation in C. perfringens may have evolved from the bacterial chromosome segregation system and, accordingly, suggest the possibility that double-stranded DNA is transferred through the Tcp conjugation apparatus.

AB - Conjugative DNA transfer is a major factor in the dissemination of antibiotic resistance and virulence genes. In the Gram-positive pathogen Clostridium perfringens, the majority of conjugative plasmids share the conserved tcp locus that governs the assembly of the transfer system. Here, we describe multiple structures of the coupling protein TcpA, an essential ATPase that is suggested to provide the mechanical force to propel the DNA through the transfer apparatus. The structures of TcpA in the presence and absence of nucleotides revealed conformational rearrangements and highlight a crucial role for the unstructured C terminus. Our findings reveal that TcpA shares most structural similarity with the FtsK DNA translocase, a central component of the bacterial cell division machinery. Our structural data suggest that conjugation in C. perfringens may have evolved from the bacterial chromosome segregation system and, accordingly, suggest the possibility that double-stranded DNA is transferred through the Tcp conjugation apparatus.

KW - ATPase

KW - Clostridium perfringens

KW - conjugation

KW - coupling protein

KW - FtsK

KW - pCW3

KW - TcpA

KW - translocase

UR - http://www.scopus.com/inward/record.url?scp=85150305503&partnerID=8YFLogxK

U2 - 10.1016/j.str.2023.02.001

DO - 10.1016/j.str.2023.02.001

M3 - Article

C2 - 36841236

AN - SCOPUS:85150305503

SN - 0969-2126

VL - 31

SP - 455-463.e4

JO - Structure

JF - Structure

IS - 4

ER -

TcpA from the Clostridium perfringens plasmid pCW3 is more closely related to the DNA translocase FtsK than to coupling proteins

Abstract

Keywords

Access to Document

Other files and links

Molecular basis for conjugative transfer of antibiotic resistance genes in Gram positive pathogens

Australian Synchrotron

Macromolecular Crystallisation Facility

Monash Proteomics & Metabolomics Facility

Cite this

TcpA from the Clostridium perfringens plasmid pCW3 is more closely related to the DNA translocase FtsK than to coupling proteins

Abstract

Keywords

Access to Document

Other files and links

Projects

Molecular basis for conjugative transfer of antibiotic resistance genes in Gram positive pathogens

Equipment

Australian Synchrotron

Macromolecular Crystallisation Facility

Monash Proteomics & Metabolomics Facility

Cite this