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

Jack Alfred 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; C. Icke; A. Teo; S-S Chng; David I Roper; T. Lithgow; A. F. Cunningham; M. Banzhaf; M. Overduin; I. R. Henderson

doi:10.1101/2020.08.10.244616

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

Jack Alfred 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, C. IckeA. Teo, S-S Chng, David I Roper, T. Lithgow, A. F. Cunningham, M. Banzhaf, M. Overduin, I. R. Henderson

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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 DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds to anionic phospholipids through an extensive membrane:protein interface providing evidence of subcellular localization of these phospholipids within the outer membrane. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope. ### Competing Interest Statement The authors have declared no competing interest.

Original language	English
DOIs	https://doi.org/10.1101/2020.08.10.244616
Publication status	Published - 10 Aug 2020

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.

Keywords

biochemistry

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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. (2020). Structure-function analyses of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localisation. https://doi.org/10.1101/2020.08.10.244616

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

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 DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds to anionic phospholipids through an extensive membrane:protein interface providing evidence of subcellular localization of these phospholipids within the outer membrane. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope. ### Competing Interest Statement The authors have declared no competing interest.",

keywords = "biochemistry",

author = "Bryant, {Jack Alfred} 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 Z-S Chong and C. Icke and A. Teo and S-S Chng and Roper, {David I} and T. Lithgow and Cunningham, {A. F.} and M. Banzhaf and M. Overduin and Henderson, {I. R.}",

note = "The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.",

year = "2020",

month = aug,

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doi = "10.1101/2020.08.10.244616",

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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, Z-S, Icke, C, Teo, A, Chng, S-S, Roper, DI, Lithgow, T, Cunningham, AF, Banzhaf, M, Overduin, M & Henderson, IR 2020 'Structure-function analyses of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localisation'. https://doi.org/10.1101/2020.08.10.244616

TY - UNPB

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

AU - Bryant, Jack Alfred

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 - Icke, C.

AU - Teo, A.

AU - Chng, S-S

AU - Roper, David I

AU - Lithgow, T.

AU - Cunningham, A. F.

AU - Banzhaf, M.

AU - Overduin, M.

AU - Henderson, I. R.

N1 - The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.

PY - 2020/8/10

Y1 - 2020/8/10

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 DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds to anionic phospholipids through an extensive membrane:protein interface providing evidence of subcellular localization of these phospholipids within the outer membrane. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope. ### Competing Interest Statement The authors have declared no competing interest.

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 DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds to anionic phospholipids through an extensive membrane:protein interface providing evidence of subcellular localization of these phospholipids within the outer membrane. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope. ### Competing Interest Statement The authors have declared no competing interest.

KW - biochemistry

UR - https://www.biorxiv.org/content/10.1101/2020.08.10.244616v1

U2 - 10.1101/2020.08.10.244616

DO - 10.1101/2020.08.10.244616

M3 - Preprint

BT - Structure-function analyses of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localisation

ER -

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

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