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A Prokaryotic Membrane Sculpting BAR Domain Protein

Phillips, Daniel A. and Zacharoff, Lori A. and Hampton, Cheri M. and Chong, Grace W. and Malanoski, Anthony P. and Metskas, Lauren Ann and Xu, Shuai and Bird, Lina J. and Eddie, Brian J. and Jensen, Grant J. and Drummy, Lawrence F. and El-Naggar, Mohamed Y. and Glaven, Sarah M. (2020) A Prokaryotic Membrane Sculpting BAR Domain Protein. . (Unpublished) https://resolver.caltech.edu/CaltechAUTHORS:20200203-093321992

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Abstract

Bin/Amphiphysin/RVS (BAR) domain proteins belong to a superfamily of coiled-coil proteins influencing membrane curvature in eukaryotes and are associated with vesicle biogenesis, vesicle-mediated protein trafficking, and intracellular signaling. Here we report the first prokaryotic BAR domain protein, BdpA, from Shewanella oneidensis MR-1, known to produce redox-active membrane vesicles and micrometer-scale outer membrane extensions (OMEs). BdpA is required for uniform size distribution of membrane vesicles and scaffolding OMEs into a consistent diameter and curvature. Cryogenic transmission electron microscopy reveals a strain lacking BdpA produces lobed, disordered OMEs rather than membrane tubes produced by the wild type strain. Overexpression of BdpA promotes OME formation during conditions where they are less common. Heterologous expression results in OME production in Marinobacter atlanticus and Escherichia coli. Based on the ability of BdpA to alter membrane curvature in vivo, we propose that BdpA and its homologs comprise a newly identified class of prokaryotic BAR (P-BAR) domains.


Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/2020.01.30.926147DOIDiscussion Paper
ORCID:
AuthorORCID
Zacharoff, Lori A.0000-0001-8657-0968
Hampton, Cheri M.0000-0003-0069-8712
Malanoski, Anthony P.0000-0001-6192-888X
Metskas, Lauren Ann0000-0002-8073-6960
Bird, Lina J.0000-0003-4127-4756
Eddie, Brian J.0000-0002-3559-3892
Jensen, Grant J.0000-0003-1556-4864
Drummy, Lawrence F.0000-0002-6452-5768
Glaven, Sarah M.0000-0003-0857-3391
Additional Information: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. bioRxiv preprint first posted online Jan. 31, 2020. We thank Dr. Jeffery Gralnick for helpful discussions and advice; Dr. Adam Meyer and Dr. Chris Voigt for the DAPG-inducible Marionette promoter; Dr. Annie Moradian and Dr. Mike Sweredoski and the California Institute of Technology Proteome Exploration Lab for useful discussions on the preparation and analysis of proteomics data. Some of the cryo-TEM work was done in the Beckman Institute Resource Center for Transmission Electron Microscopy at Caltech. This work was supported by the United States Department of Defense Synthetic Biology for Military Environments (SBME) Applied Research for the Advancement of Science and Technology Priorities (ARAP) program. Work in ME-N's lab was supported by the U.S. Office of Naval Research Multidisciplinary University Research Initiative Grant No. N00014-18-1-2632. LAZ was partially supported by the National Science Foundation grant DEB-1542527. SX was supported by the Division of Chemical Sciences, Geosciences, and Biosciences, Office of Basic Energy Sciences of the U.S. Department of Energy through grant DE-FG02-13ER16415. Work in GJJ’s lab was supported by the National Institute of Health (GM122588 to GJJ). Author Contributions: DP and LZ conceived the study independently then combined projects when complementary data on BdpA was discovered. LZ purified OMVs, prepared samples for LC MS-MS, and performed DLS measurements. LZ and SX made electrochemical measurements and analysis. DP conducted BdpA domain prediction and validation analysis, generated the p452-bdpA plasmid, ΔbdpA and p452-bdpA strains. DP and GC conducted fluorescence imaging experiments, and DP, LZ, and GC analyzed the data. LB adapted the Marionette sensor (PphlF-YFP) into pBBR1-mcs2. LZ and LAM performed cryo-TEM of OMVs. GC and LZ analyzed perfusion flow system data. CH, DP, and LD performed cryo-TEM experiments of OMEs and image processing / analysis. DP and AM generated phylogenetic data, and DP, AM, and BE analyzed the data. DP, LZ, CM, GC, AM, LAM, BE, GJJ, LD, MEN, and SG provided data interpretation. DP, LZ, MEN, and SG wrote the manuscript, with input from all coauthors.
Funders:
Funding AgencyGrant Number
Office of Naval Research (ONR)N00014-18-1-2632
NSFDEB-1542527
Department of Energy (DOE)DE-FG02-13ER16415
NIHGM122588
Record Number:CaltechAUTHORS:20200203-093321992
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20200203-093321992
Official Citation:A Prokaryotic Membrane Sculpting BAR Domain Protein. Daniel A. Phillips, Lori A. Zacharoff, Cheri M. Hampton, Grace W. Chong, Anthony P. Malanoski, Lauren Ann Metskas, Shuai Xu, Lina J. Bird, Brian J. Eddie, Grant J. Jensen, Lawrence F. Drummy, Mohamed Y. El-Naggar, Sarah M. Glaven. bioRxiv 2020.01.30.926147; doi: https://doi.org/10.1101/2020.01.30.926147
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:101068
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:03 Feb 2020 17:47
Last Modified:12 Jun 2020 17:12

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