Teasing apart the evolution of lipoprotein trafficking in gram-negative bacteria reveals a bifunctional LolA

• Type of Material: Article
• Authors: Hannah C Smith, Emory University; Kerrie L May, Emory University; Marcin Grabowicz, Emory University
• Language: English
• Date: 2023-02-07
• Publisher: PNAS
• Publication Version: Final Published Version
• Copyright Statement: © The Author(s) 2023
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1073/pnas.2218473120
• Title of Journal or Parent Work: Proceedings of the National Academy of Sciences of the United States of America
• Volume: 120
• Issue: 6
• Start Page: e2218473120
• End Page: e2218473120
• Keywords: Molecular Chaperones; Bacterial Outer Membrane Proteins; outer membrane assembly; lipoprotein trafficking; LolA; Escherichia coli; Gram-Negative Bacteria; Lipoproteins; Escherichia coli Proteins; Periplasmic Binding Proteins; Caulobacter vibrioides

Abstract

The outer membrane (OM) is the defining feature of gram-negative bacteria and is an essential organelle. Accordingly, OM assembly pathways and their essential protein components are conserved throughout all gram-negative species. Lipoprotein trafficking lies at the heart of OM assembly since it supplies several different biogenesis machines with essential lipoproteins. The Escherichia coli Lol trafficking pathway relies on an inner membrane LolCDE transporter that transfers newly made lipoproteins to the chaperone LolA, which rapidly traffics lipoproteins across the periplasm to LolB for insertion into the OM. Strikingly, many gram-negative species (like Caulobacter vibrioides) do not produce LolB, yet essential lipoproteins are still trafficked to the OM. How the final step of trafficking occurs in these organisms has remained a long-standing mystery. We demonstrate that LolA from C. vibrioides can complement the deletion of both LolA and LolB in E. coli, revealing that this protein possesses both chaperone and insertion activities. Moreover, we define the region of C. vibrioides LolA that is responsible for its bifunctionality. This knowledge enabled us to convert E. coli LolA into a similarly bifunctional protein, capable of chaperone and insertion activities. We propose that a bifunctional LolA eliminates the need for LolB. Our findings provide an explanation for why some gram-negative species have retained an essential LolA yet completely lack a dedicated LolB protein.