Conserved features in TamA enable interaction with TamB to drive the activity of the translocation and assembly module

Joel Pearson Selkrig, Matthew J Belousoff, Stephen J Headey, Eva Heinz, Takuya Shiota, Hsin-Hui Shen, Simone A Beckham, Rebecca S Bamert, Minh-Duy Phan, Mark A Schembri, Matthew C Wilce, Martin J Scanlon, Richard A Strugnell, Trevor James Lithgow

Research output: Contribution to journalArticleResearchpeer-review

19 Citations (Scopus)

Abstract

The biogenesis of membranes from constituent proteins and lipids is a fundamental aspect of cell biology. In the case of proteins assembled into bacterial outer membranes, an overarching question concerns how the energy required for protein insertion and folding is accessed at this remote location of the cell. The translocation and assembly module (TAM) is a nanomachine that functions in outer membrane biogenesis and virulence in diverse bacterial pathogens. Here we demonstrate the interactions through which TamA and TamB subunits dock to bridge the periplasm, and unite the outer membrane aspects to the inner membrane of the bacterial cell. We show that specific functional features in TamA have been conserved through evolution, including residues surrounding the lateral gate and an extensive surface of the POTRA domains. Analysis by nuclear magnetic resonance spectroscopy and small angle X-ray scattering document the characteristic structural features of these POTRA domains and demonstrate rigidity in solution. Quartz crystal microbalance measurements pinpoint which POTRA domain specifically docks the TamB subunit of the nanomachine. We speculate that the POTRA domain of TamA functions as a lever arm in order to drive the activity of the TAM, assembling proteins into bacterial outer membranes.
Original languageEnglish
Article number12905
Number of pages12
JournalScientific Reports
Volume5
DOIs
Publication statusPublished - 2015

Cite this

Selkrig, Joel Pearson ; Belousoff, Matthew J ; Headey, Stephen J ; Heinz, Eva ; Shiota, Takuya ; Shen, Hsin-Hui ; Beckham, Simone A ; Bamert, Rebecca S ; Phan, Minh-Duy ; Schembri, Mark A ; Wilce, Matthew C ; Scanlon, Martin J ; Strugnell, Richard A ; Lithgow, Trevor James. / Conserved features in TamA enable interaction with TamB to drive the activity of the translocation and assembly module. In: Scientific Reports. 2015 ; Vol. 5.
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title = "Conserved features in TamA enable interaction with TamB to drive the activity of the translocation and assembly module",
abstract = "The biogenesis of membranes from constituent proteins and lipids is a fundamental aspect of cell biology. In the case of proteins assembled into bacterial outer membranes, an overarching question concerns how the energy required for protein insertion and folding is accessed at this remote location of the cell. The translocation and assembly module (TAM) is a nanomachine that functions in outer membrane biogenesis and virulence in diverse bacterial pathogens. Here we demonstrate the interactions through which TamA and TamB subunits dock to bridge the periplasm, and unite the outer membrane aspects to the inner membrane of the bacterial cell. We show that specific functional features in TamA have been conserved through evolution, including residues surrounding the lateral gate and an extensive surface of the POTRA domains. Analysis by nuclear magnetic resonance spectroscopy and small angle X-ray scattering document the characteristic structural features of these POTRA domains and demonstrate rigidity in solution. Quartz crystal microbalance measurements pinpoint which POTRA domain specifically docks the TamB subunit of the nanomachine. We speculate that the POTRA domain of TamA functions as a lever arm in order to drive the activity of the TAM, assembling proteins into bacterial outer membranes.",
author = "Selkrig, {Joel Pearson} and Belousoff, {Matthew J} and Headey, {Stephen J} and Eva Heinz and Takuya Shiota and Hsin-Hui Shen and Beckham, {Simone A} and Bamert, {Rebecca S} and Minh-Duy Phan and Schembri, {Mark A} and Wilce, {Matthew C} and Scanlon, {Martin J} and Strugnell, {Richard A} and Lithgow, {Trevor James}",
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Conserved features in TamA enable interaction with TamB to drive the activity of the translocation and assembly module. / Selkrig, Joel Pearson; Belousoff, Matthew J; Headey, Stephen J; Heinz, Eva; Shiota, Takuya; Shen, Hsin-Hui; Beckham, Simone A; Bamert, Rebecca S; Phan, Minh-Duy; Schembri, Mark A; Wilce, Matthew C; Scanlon, Martin J; Strugnell, Richard A; Lithgow, Trevor James.

In: Scientific Reports, Vol. 5, 12905, 2015.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Conserved features in TamA enable interaction with TamB to drive the activity of the translocation and assembly module

AU - Selkrig, Joel Pearson

AU - Belousoff, Matthew J

AU - Headey, Stephen J

AU - Heinz, Eva

AU - Shiota, Takuya

AU - Shen, Hsin-Hui

AU - Beckham, Simone A

AU - Bamert, Rebecca S

AU - Phan, Minh-Duy

AU - Schembri, Mark A

AU - Wilce, Matthew C

AU - Scanlon, Martin J

AU - Strugnell, Richard A

AU - Lithgow, Trevor James

PY - 2015

Y1 - 2015

N2 - The biogenesis of membranes from constituent proteins and lipids is a fundamental aspect of cell biology. In the case of proteins assembled into bacterial outer membranes, an overarching question concerns how the energy required for protein insertion and folding is accessed at this remote location of the cell. The translocation and assembly module (TAM) is a nanomachine that functions in outer membrane biogenesis and virulence in diverse bacterial pathogens. Here we demonstrate the interactions through which TamA and TamB subunits dock to bridge the periplasm, and unite the outer membrane aspects to the inner membrane of the bacterial cell. We show that specific functional features in TamA have been conserved through evolution, including residues surrounding the lateral gate and an extensive surface of the POTRA domains. Analysis by nuclear magnetic resonance spectroscopy and small angle X-ray scattering document the characteristic structural features of these POTRA domains and demonstrate rigidity in solution. Quartz crystal microbalance measurements pinpoint which POTRA domain specifically docks the TamB subunit of the nanomachine. We speculate that the POTRA domain of TamA functions as a lever arm in order to drive the activity of the TAM, assembling proteins into bacterial outer membranes.

AB - The biogenesis of membranes from constituent proteins and lipids is a fundamental aspect of cell biology. In the case of proteins assembled into bacterial outer membranes, an overarching question concerns how the energy required for protein insertion and folding is accessed at this remote location of the cell. The translocation and assembly module (TAM) is a nanomachine that functions in outer membrane biogenesis and virulence in diverse bacterial pathogens. Here we demonstrate the interactions through which TamA and TamB subunits dock to bridge the periplasm, and unite the outer membrane aspects to the inner membrane of the bacterial cell. We show that specific functional features in TamA have been conserved through evolution, including residues surrounding the lateral gate and an extensive surface of the POTRA domains. Analysis by nuclear magnetic resonance spectroscopy and small angle X-ray scattering document the characteristic structural features of these POTRA domains and demonstrate rigidity in solution. Quartz crystal microbalance measurements pinpoint which POTRA domain specifically docks the TamB subunit of the nanomachine. We speculate that the POTRA domain of TamA functions as a lever arm in order to drive the activity of the TAM, assembling proteins into bacterial outer membranes.

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