Self-assembling Shell Proteins PduA and PduJ have Essential and Redundant Roles in Bacterial Microcompartment Assembly

Kennedy, Nolan W.; Ikonomova, Svetlana P.; Slininger Lee, Marilyn; Raeder, Henry W.; Tullman-Ercek, Danielle

doi:10.1016/j.jmb.2020.11.020

Self-assembling Shell Proteins PduA and PduJ have Essential and Redundant Roles in Bacterial Microcompartment Assembly

Journal Article · Sat Nov 21 00:00:00 EST 2020 · Journal of Molecular Biology

DOI:https://doi.org/10.1016/j.jmb.2020.11.020· OSTI ID:1853101

Kennedy, Nolan W. ^[1]; Ikonomova, Svetlana P. ^[2]; Slininger Lee, Marilyn ^[3]; Raeder, Henry W. ^[4]; Tullman-Ercek, Danielle ^[5]

Northwestern Univ., Evanston, IL (United States). Interdisciplinary Biological Sciences Graduate Program; Northwestern Univ., Evanston, IL (United States)
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

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.

View Accepted Manuscript (DOE)

Research Organization:: Northwestern Univ., Evanston, IL (United States)

Sponsoring Organization:: National Institutes of Health (NIH); National Science Foundation (NSF); US Army Research Office (ARO); USDOE; USDOE Office of Science (SC)

Grant/Contract Number:: SC0019337

OSTI ID:: 1853101

Alternate ID(s):: OSTI ID: 1775967

Journal Information:: Journal of Molecular Biology, Journal Name: Journal of Molecular Biology Journal Issue: 2 Vol. 433; ISSN 0022-2836

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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