Functions of the BamBCDE lipoproteins revealed by bypass mutations in BamA

Elizabeth M. Hart, Thomas J. Silhavy

Research output: Contribution to journalArticlepeer-review

Abstract

The heteropentomeric β-barrel assembly machine (BAM complex) is responsible for folding and inserting a diverse array of β-barrel outer membrane proteins (OMPs) into the outer membrane (OM) of Gram-negative bacteria. The BAM complex contains two essential proteins, the β-barrel OMP BamA and a lipoprotein BamD, whereas the auxiliary lipoproteins BamBCE are individually nonessential. Here, we identify and characterize three bamA mutations, the E-to-K change at position 470 (bamAE470K), the A-to-P change at position 496 (bamAA496P), and the A-to-S change at position 499 (bamAA499S), that suppress the otherwise lethal ΔbamD, ΔbamB ΔbamC ΔbamE, and ΔbamC ΔbamD ΔbamE mutations. The viability of cells lacking different combinations of BAM complex lipoproteins provides the opportunity to examine the role of the individual proteins in OMP assembly. Results show that, in wild-type cells, BamBCE share a redundant function; at least one of these lipoproteins must be present to allow BamD to coordinate productively with BamA. Besides BamA regulation, BamD shares an additional essential function that is redundant with a second function of BamB. Remarkably, bamAE470K suppresses both, allowing the construction of a BAM complex composed solely of BamAE470K that is able to assemble OMPs in the absence of BamBCDE. This work demonstrates that the BAM complex lipoproteins do not participate in the catalytic folding of OMP substrates but rather function to increase the efficiency of the assembly process by coordinating and regulating the assembly of diverse OMP substrates.

Original languageEnglish (US)
Article numbere00401-20
JournalJournal of bacteriology
Volume202
Issue number21
DOIs
StatePublished - Oct 8 2020

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Keywords

  • BAM complex
  • Escherichia coli
  • Gram-negative bacteria
  • Outer membrane
  • Outer membrane biogenesis

Fingerprint Dive into the research topics of 'Functions of the BamBCDE lipoproteins revealed by bypass mutations in BamA'. Together they form a unique fingerprint.

Cite this