Structure of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localization

J. A. Bryant; F. C. Morris; T. J. Knowles; R. Maderbocus; E. Heinz; G. Boelter; D. Alodaini; A. Colyer; P. J. Wotherspoon; K. A. Staunton; M. Jeeves; D. F. Browning; Y. R. Sevastsyanovich; T. J. Wells; A. E. Rossiter; V. N. Bavro; P. Sridhar; D. G. Ward; Z. S. Chong; E. C.A. Goodall; C. Icke; A. Teo; S. S. Chng; D. I. Roper; T. Lithgow; A. F. Cunningham; M. Banzhaf; M. Overduin; I. R. Henderson

doi:10.7554/eLife.62614

Structure of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localization

J. A. Bryant, F. C. Morris, T. J. Knowles, R. Maderbocus, E. Heinz, G. Boelter, D. Alodaini, A. Colyer, P. J. Wotherspoon, K. A. Staunton, M. Jeeves, D. F. Browning, Y. R. Sevastsyanovich, T. J. Wells, A. E. Rossiter, V. N. Bavro, P. Sridhar, D. G. Ward, Z. S. Chong, E. C.A. GoodallC. Icke, A. Teo, S. S. Chng, D. I. Roper, T. Lithgow, A. F. Cunningham, M. Banzhaf, M. Overduin, I. R. Henderson^*

^*Corresponding author for this work

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

25 Citations (Scopus)

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Abstract

The Gram-negative outer membrane envelops the bacterium and functions as a permeability barrier against antibiotics, detergents and environmental stresses. Some virulence factors serve to maintain the integrity of the outer membrane, including DolP (formerly YraP) a protein of unresolved structure and function. Here we reveal DolP is a lipoprotein functionally conserved among Gram-negative bacteria and that loss of DolP increases membrane fluidity. We present the NMR solution structure for Escherichia coli DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds anionic phospholipids through an extensive membrane:protein interface. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site, providing evidence of subcellular localization of these phospholipids within the outer membrane. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope.

Original language	English
Article number	e62614
Number of pages	61
Journal	eLife
Volume	9
Early online date	14 Dec 2020
DOIs	https://doi.org/10.7554/eLife.62614
Publication status	Published - 13 Jan 2021
Externally published	Yes

Funding

This work was supported by the BBSRC (I.R.H. and M.O.: BB/M00810X/1 and BB/L00335X/1; T.J.K. BB/P009840/1), NSERC RGPIN-2018-04994 and Campus Alberta Innovation Program (RCP-12-002C) (M.O.). We would like to thank Georgia L. Isom and Catherine A. Wardius for technical assistance in the laboratory. We thank Professor Corinne Spickett for use of mass spectrometry facilities for phospholipid analyses. We also thank Professor Jeff Cole for critical advice in development of the project.

Keywords

antibiotics
membrane:protein interface
bacterial cell envelope

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10.7554/eLife.62614Licence: CC BY 4.0

Bryant-etal-elife-Structure-of-dual-BON-domain-protein-DolP-identifies-phospholipid-bindingFinal published version, 3.3 MBLicence: CC BY 4.0

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Bryant, J. A., Morris, F. C., Knowles, T. J., Maderbocus, R., Heinz, E., Boelter, G., Alodaini, D., Colyer, A., Wotherspoon, P. J., Staunton, K. A., Jeeves, M., Browning, D. F., Sevastsyanovich, Y. R., Wells, T. J., Rossiter, A. E., Bavro, V. N., Sridhar, P., Ward, D. G., Chong, Z. S., ... Henderson, I. R. (2021). Structure of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localization. eLife, 9, Article e62614. https://doi.org/10.7554/eLife.62614

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title = "Structure of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localization",

abstract = "The Gram-negative outer membrane envelops the bacterium and functions as a permeability barrier against antibiotics, detergents and environmental stresses. Some virulence factors serve to maintain the integrity of the outer membrane, including DolP (formerly YraP) a protein of unresolved structure and function. Here we reveal DolP is a lipoprotein functionally conserved among Gram-negative bacteria and that loss of DolP increases membrane fluidity. We present the NMR solution structure for Escherichia coli DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds anionic phospholipids through an extensive membrane:protein interface. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site, providing evidence of subcellular localization of these phospholipids within the outer membrane. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope.",

keywords = "antibiotics, membrane:protein interface, bacterial cell envelope",

author = "Bryant, \{J. A.\} and Morris, \{F. C.\} and Knowles, \{T. J.\} and R. Maderbocus and E. Heinz and G. Boelter and D. Alodaini and A. Colyer and Wotherspoon, \{P. J.\} and Staunton, \{K. A.\} and M. Jeeves and Browning, \{D. F.\} and Sevastsyanovich, \{Y. R.\} and Wells, \{T. J.\} and Rossiter, \{A. E.\} and Bavro, \{V. N.\} and P. Sridhar and Ward, \{D. G.\} and Chong, \{Z. S.\} and Goodall, \{E. C.A.\} and C. Icke and A. Teo and Chng, \{S. S.\} and Roper, \{D. I.\} and T. Lithgow and Cunningham, \{A. F.\} and M. Banzhaf and M. Overduin and Henderson, \{I. R.\}",

year = "2021",

month = jan,

day = "13",

doi = "10.7554/eLife.62614",

language = "English",

volume = "9",

journal = "eLife",

issn = "2050-084X",

publisher = "eLife Sciences Publications Ltd",

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Bryant, JA, Morris, FC, Knowles, TJ, Maderbocus, R, Heinz, E, Boelter, G, Alodaini, D, Colyer, A, Wotherspoon, PJ, Staunton, KA, Jeeves, M, Browning, DF, Sevastsyanovich, YR, Wells, TJ, Rossiter, AE, Bavro, VN, Sridhar, P, Ward, DG, Chong, ZS, Goodall, ECA, Icke, C, Teo, A, Chng, SS, Roper, DI, Lithgow, T, Cunningham, AF, Banzhaf, M, Overduin, M & Henderson, IR 2021, 'Structure of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localization', eLife, vol. 9, e62614. https://doi.org/10.7554/eLife.62614

TY - JOUR

T1 - Structure of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localization

AU - Bryant, J. A.

AU - Morris, F. C.

AU - Knowles, T. J.

AU - Maderbocus, R.

AU - Heinz, E.

AU - Boelter, G.

AU - Alodaini, D.

AU - Colyer, A.

AU - Wotherspoon, P. J.

AU - Staunton, K. A.

AU - Jeeves, M.

AU - Browning, D. F.

AU - Sevastsyanovich, Y. R.

AU - Wells, T. J.

AU - Rossiter, A. E.

AU - Bavro, V. N.

AU - Sridhar, P.

AU - Ward, D. G.

AU - Chong, Z. S.

AU - Goodall, E. C.A.

AU - Icke, C.

AU - Teo, A.

AU - Chng, S. S.

AU - Roper, D. I.

AU - Lithgow, T.

AU - Cunningham, A. F.

AU - Banzhaf, M.

AU - Overduin, M.

AU - Henderson, I. R.

PY - 2021/1/13

Y1 - 2021/1/13

N2 - The Gram-negative outer membrane envelops the bacterium and functions as a permeability barrier against antibiotics, detergents and environmental stresses. Some virulence factors serve to maintain the integrity of the outer membrane, including DolP (formerly YraP) a protein of unresolved structure and function. Here we reveal DolP is a lipoprotein functionally conserved among Gram-negative bacteria and that loss of DolP increases membrane fluidity. We present the NMR solution structure for Escherichia coli DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds anionic phospholipids through an extensive membrane:protein interface. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site, providing evidence of subcellular localization of these phospholipids within the outer membrane. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope.

AB - The Gram-negative outer membrane envelops the bacterium and functions as a permeability barrier against antibiotics, detergents and environmental stresses. Some virulence factors serve to maintain the integrity of the outer membrane, including DolP (formerly YraP) a protein of unresolved structure and function. Here we reveal DolP is a lipoprotein functionally conserved among Gram-negative bacteria and that loss of DolP increases membrane fluidity. We present the NMR solution structure for Escherichia coli DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds anionic phospholipids through an extensive membrane:protein interface. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site, providing evidence of subcellular localization of these phospholipids within the outer membrane. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope.

KW - antibiotics

KW - membrane:protein interface

KW - bacterial cell envelope

UR - https://www.scopus.com/pages/publications/85100069579

U2 - 10.7554/eLife.62614

DO - 10.7554/eLife.62614

M3 - Article

C2 - 33315009

AN - SCOPUS:85100069579

SN - 2050-084X

VL - 9

JO - eLife

JF - eLife

M1 - e62614

ER -

Structure of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localization

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