Genetic analysis of the protein shell of the microcompartments involved in coenzyme B12-dependent 1,2-propanediol degradation by Salmonella.
Hundreds of bacterial species use microcompartments (MCPs) to optimize metabolic pathways that have toxic or volatile intermediates. MCPs consist of a protein shell encapsulating specific metabolic enzymes. In Salmonella, an MCP is used for 1,2-propanediol utilization (Pdu MCP). The shell of this MCP is composed of eight different types of polypeptides, but their specific functions are uncertain. Here, we individually deleted the eight genes encoding the shell proteins of the Pdu MCP. The effects of each mutation on 1,2-PD degradation and MCP structure were determined by electron microscopy and growth studies. Deletion of the pduBB', pduJ, or pduN gene severely impaired MCP formation, and the observed defects were consistent with roles as facet, edge, or vertex protein, respectively. Metabolite measurements showed that pduA, pduBB', pduJ, or pduN deletion mutants accumulated propionaldehyde to toxic levels during 1,2-PD catabolism, indicating that the integrity of the shell was disrupted. Deletion of the pduK, pduT, or pduU gene did not substantially affect MCP structure or propionaldehyde accumulation, suggesting they are nonessential to MCP formation. However, the pduU or pduT deletion mutants grew more slowly than the wild type on 1,2-PD at saturating B(12), indicating that they are needed for maximal activity of the 1,2-PD degradative enzymes encased within the MCP shell. Considering recent crystallography studies, this suggests that PduT and PduU may mediate the transport of enzyme substrates/cofactors across the MCP shell. Interestingly, a pduK deletion caused MCP aggregation, suggesting a role in the spatial organization of MCP within the cytoplasm or perhaps in segregation at cell division.
Iowa State University, Department of Biochemistry, Biophysics and Molecular Biology, Ames, IA 50011, USA.
Shouqiang
S
Cheng
Sharmistha
S
Sinha
Chenguang
C
Fan
Yu
Y
Liu
Thomas A
TA
Bobik
D000447:Aldehydes / Q000378:metabolism / Q000633:toxicity; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D003038:Cobamides / Q000378:metabolism*; D017353:Gene Deletion; D008854:Microscopy, Electron; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure; D019946:Propylene Glycol / Q000378:metabolism*; D012486:Salmonella typhimurium / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure
D000447:Aldehydes; D001426:Bacterial Proteins; D003038:Cobamides; D019946:Propylene Glycol; C005556:propionaldehyde; C000913:cobamamide
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