Abstract
Gram-negative bacteria employ the Type II Secretion System (T2SS) to not only secrete an array of soluble effectors such as toxins to the extracellular space, but also to facilitate the surface localization of enzymes and adhesins that are beneficial to life in different environments. For example, the pullulan degrading enzyme pullulanase (PulA) from Klebsiella pneumoniae and the recently discovered adhesin InvL from Acinetobacter baumannii are initially expressed with a lipobox containing signal peptide, resulting in their N-terminal acylation and subsequent surface anchoring after T2SS mediated export. While outer membrane translocation of both soluble and surface associated T2SS effectors depends on the T2SS secretin GspD, it is unclear how lipoproteins are accommodated by the T2SS during transport to the cell surface. Here, we identify a role for GspN in the outer membrane translocation of InvL in the opportunistic pathogen A. baumannii. Additional putative lipoproteins are found to have a similar GspN dependence for secretion, while soluble proteins are secreted independently of GspN. We demonstrate that a specific sorting motif C-terminal to the lipobox is required for GspN-dependent surface localization. Based on structural predictions, GspN belongs to the larger AsmA-like protein family that includes both eukaryotic and prokaryotic members. This protein family has been implicated in phospholipid transport, but here we expand the role for this family to include transport of lipoproteins. We also confirm that the GspN homolog PulN is required for PulA surface localization in K. pneumoniae.