Self-assembling Shell Proteins PduA and PduJ have Essential and Redundant Roles in Bacterial Microcompartment Assembly
Abstract
Protein self-assembly is a common and essential biological phenomenon, and bacterial microcompartments present a promising model system to study this process. Bacterial microcompartments are large, protein-based organelles which natively carry out processes important for carbon fixation in cyanobacteria and the survival of enteric bacteria. These structures are increasingly popular with biological engineers due to their potential utility as nanobioreactors or drug delivery vehicles. However, the limited understanding of the assembly mechanism of these bacterial microcompartments hinders efforts to repurpose them for non-native functions. Here, we comprehensively investigate proteins involved in the assembly of the 1,2-propanediol utilization bacterial microcompartment from Salmonella enterica serovar Typhimurium LT2, one of the most widely studied microcompartment systems. We first demonstrate that two shell proteins, PduA and PduJ, have a high propensity for self-assembly upon overexpression, and we provide a novel method for self-assembly quantification. Using genomic knock-outs and knock-ins, we systematically show that these two proteins play an essential and redundant role in bacterial microcompartment assembly that cannot be compensated by other shell proteins. At least one of the two proteins PduA and PduJ must be present for the bacterial microcompartment shell to assemble. We also demonstrate that assembly-deficient variants of these proteins are unablemore »
- Authors:
-
- Northwestern Univ., Evanston, IL (United States). Interdisciplinary Biological Sciences Graduate Program
- Northwestern Univ., Evanston, IL (United States). Dept. of Chemical and Biological Engineering
- Northwestern Univ., Evanston, IL (United States). Dept. of Chemical and Biological Engineering; US Army Combat Capabilities Development Command Chemical Biological Center, Edgewood, MD (United States)
- Northwestern Univ., Evanston, IL (United States).Weinberg College of Arts and Sciences. Molecular Biosciences Program
- Northwestern Univ., Evanston, IL (United States). Dept. of Chemical and Biological Engineering; Northwestern Univ., Evanston, IL (United States). Center for Synthetic Biology
- Publication Date:
- Research Org.:
- Northwestern Univ., Evanston, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC); National Science Foundation (NSF); US Army Research Office (ARO); National Institutes of Health (NIH)
- OSTI Identifier:
- 1853101
- Alternate Identifier(s):
- OSTI ID: 1775967
- Grant/Contract Number:
- SC0019337; MCB1150567; W911NF-19-1-0298; DGE-1842165; DMR-1720139; ECCS-1542205; T32GM008449
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Molecular Biology
- Additional Journal Information:
- Journal Volume: 433; Journal Issue: 2; Journal ID: ISSN 0022-2836
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 1,2-propanediol utilization MCP; gene knockout; oligomerization; Salmonella enterica serovar Typhimurium LT2; rapid self-assembly assay
Citation Formats
Kennedy, Nolan W., Ikonomova, Svetlana P., Slininger Lee, Marilyn, Raeder, Henry W., and Tullman-Ercek, Danielle. Self-assembling Shell Proteins PduA and PduJ have Essential and Redundant Roles in Bacterial Microcompartment Assembly. United States: N. p., 2020.
Web. doi:10.1016/j.jmb.2020.11.020.
Kennedy, Nolan W., Ikonomova, Svetlana P., Slininger Lee, Marilyn, Raeder, Henry W., & Tullman-Ercek, Danielle. Self-assembling Shell Proteins PduA and PduJ have Essential and Redundant Roles in Bacterial Microcompartment Assembly. United States. https://doi.org/10.1016/j.jmb.2020.11.020
Kennedy, Nolan W., Ikonomova, Svetlana P., Slininger Lee, Marilyn, Raeder, Henry W., and Tullman-Ercek, Danielle. Sat .
"Self-assembling Shell Proteins PduA and PduJ have Essential and Redundant Roles in Bacterial Microcompartment Assembly". United States. https://doi.org/10.1016/j.jmb.2020.11.020. https://www.osti.gov/servlets/purl/1853101.
@article{osti_1853101,
title = {Self-assembling Shell Proteins PduA and PduJ have Essential and Redundant Roles in Bacterial Microcompartment Assembly},
author = {Kennedy, Nolan W. and Ikonomova, Svetlana P. and Slininger Lee, Marilyn and Raeder, Henry W. and Tullman-Ercek, Danielle},
abstractNote = {Protein self-assembly is a common and essential biological phenomenon, and bacterial microcompartments present a promising model system to study this process. Bacterial microcompartments are large, protein-based organelles which natively carry out processes important for carbon fixation in cyanobacteria and the survival of enteric bacteria. These structures are increasingly popular with biological engineers due to their potential utility as nanobioreactors or drug delivery vehicles. However, the limited understanding of the assembly mechanism of these bacterial microcompartments hinders efforts to repurpose them for non-native functions. Here, we comprehensively investigate proteins involved in the assembly of the 1,2-propanediol utilization bacterial microcompartment from Salmonella enterica serovar Typhimurium LT2, one of the most widely studied microcompartment systems. We first demonstrate that two shell proteins, PduA and PduJ, have a high propensity for self-assembly upon overexpression, and we provide a novel method for self-assembly quantification. Using genomic knock-outs and knock-ins, we systematically show that these two proteins play an essential and redundant role in bacterial microcompartment assembly that cannot be compensated by other shell proteins. At least one of the two proteins PduA and PduJ must be present for the bacterial microcompartment shell to assemble. We also demonstrate that assembly-deficient variants of these proteins are unable to rescue microcompartment formation, highlighting the importance of this assembly property. Our work provides insight into the assembly mechanism of these bacterial organelles and will aid downstream engineering efforts.},
doi = {10.1016/j.jmb.2020.11.020},
journal = {Journal of Molecular Biology},
number = 2,
volume = 433,
place = {United States},
year = {Sat Nov 21 00:00:00 EST 2020},
month = {Sat Nov 21 00:00:00 EST 2020}
}
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