Listeria monocytogenes hijacks CD147 to ensure proper membrane protrusion formation and efficient bacterial dissemination

Abstract

Efficient cell-to-cell transfer of Listeria monocytogenes (L. monocytogenes) requires the proper formation of actin-rich membrane protrusions. To date, only the host proteins ezrin, the binding partner of ezrin, CD44, as well as cyclophilin A (CypA) have been identified as crucial components for L. monocytogenes membrane protrusion stabilization and, thus, efficient cell-to-cell movement of the microbes. Here, we examine the classical binding partner of CypA, CD147, and find that this membrane protein is also hijacked by the bacteria for their cellular dissemination. CD147 is enriched at the plasma membrane surrounding the membrane protrusions as well as the resulting invaginations generated in neighboring cells. In cells depleted of CD147, these actin-rich structures appear similar to those generated in CypA depleted cells as they are significantly shorter and more contorted as compared to their straighter counterparts formed in wild-type control cells. The presence of malformed membrane protrusions hampers the ability of L. monocytogenes to efficiently disseminate from CD147-depleted cells. Our findings uncover another important host protein needed for L. monocytogenes membrane protrusion formation and efficient microbial dissemination.

Listeria monocytogenes Exploits Host Caveolin for Cell-to-Cell Spreading

“…We recently identified CD147 as a marker of L. monocytogenes membrane invaginations (23). Thus, to quantify the frequency with which caveolin-1 delineates L. monocytogenes membrane invaginations, we coexpressed CD147-GFP with caveolin-1-mCherry in invagination-forming host cells.…”

“…From our experiments involving isolated L. monocytogenes membrane protrusions, the results implied that contact of the structures with the host cell surface alone is sufficient to trigger recruitment of caveolin-1, cavin-2, and EHD2. Coupled with our recent finding of the host plasma membrane receptor CD147 at L. monocytogenes membrane invaginations (23), the potential for the presence of a ligand(s) on the surface of protrusions, interacting with a corresponding receptor(s) on the cell forming the invaginations, presents a compelling area of further investigation. Furthermore, the visually apparent elevated levels of these caveolar proteins surrounding the structures also point to the possibility that preassembled caveolar vesicles could incorporate their proteins into the plasma membrane as an endocytic unit at sites of membrane invaginations (72,73) rather than being recruited separately.…”

“…4I and J). Similarly, we analyzed the morphology of at least 40 L. monocytogenes membrane protrusions, as the proper formation of these structures is also required for efficient bacterial cell-to-cell dissemination (23,44,53,54), and found no significant differences in the length or tortuosity of the structures (Fig. 4K and L).…”

Distribution of PDLIM1 at actin‐rich structures generated by invasive and adherent bacterial pathogens

“…Thus, to examine if PDLIM1, and/or its actin‐crosslinking binding partners (α‐actinin‐1 and palladin) are also present at L . monocytogenes membrane invaginations, we utilized a mixed‐cell assay (Dhanda, Lulic, Yu, et al, 2019; Dhanda et al, 2020). In this assay, infected cells are overlaid onto uninfected cells that had been transfected with PDLIM1‐GFP (or fluorescently‐tagged versions of its binding partners).…”

“…A common feature of L . monocytogenes and EPEC actin‐rich structures is that they are also comprised of a vast array of actin‐associated proteins such as actin nucleators (Arp2/3 complex (Kalman et al, 1999; Welch et al, 1997), formins (Fattouh et al, 2015; Velle & Campellone, 2018) and VASP (Dhanda et al, 2018; Goosney et al, 2000)), capping proteins (CapZ (David et al, 1998) and gelsolin (Goosney, DeVinney, & Finlay, 2001; Laine et al, 1998)), severing protein (ADF/cofilin (David et al, 1998; Goosney et al, 2001; Talman, Chong, Chia, Svitkina, & Agaisse, 2014)) and many crosslinking agents (palladin (Dhanda et al, 2018), α‐actinins 1 and 4 (Dabiri, Sanger, Portnoy, & Southwick, 1990; Dhanda, Lulic, Yu, et al, 2019; Finlay, Rosenshine, Donnenberg, & Kaper, 1992; Nanavati, Ashton, Sanger, & Sanger, 1994), nexilin (Law, Bonazzi, Jackson, Cossart, & Guttman, 2012), IQGAP1 (Brown, Bry, Li, & Sacks, 2008) and transgelin (Chua, Hipolito, Singerr, Solway, & Guttman, 2018)). We and others have shown actin filament crosslinkers (α‐actinin 1 (Dold, Sanger, & Sanger, 1994), palladin (Dhanda et al, 2018), nexilin (Law et al, 2012) and transgelin (Chua et al, 2018)) as key determinants of L .…”