JAM-A interacts with α3β1 integrin and tetraspanins CD151 and CD9 to regulate collective cell migration of polarized epithelial cells

Abstract

Junctional adhesion molecule (JAM)-A is a cell adhesion receptor localized at epithelial cell–cell contacts with enrichment at the tight junctions. Its role during cell–cell contact formation and epithelial barrier formation has intensively been studied. In contrast, its role during collective cell migration is largely unexplored. Here, we show that JAM-A regulates collective cell migration of polarized epithelial cells. Depletion of JAM-A in MDCK cells enhances the motility of singly migrating cells but reduces cell motility of cells embedded in a collective by impairing the dynamics of cryptic lamellipodia formation. This activity of JAM-A is observed in cells grown on laminin and collagen-I but not on fibronectin or vitronectin. Accordingly, we find that JAM-A exists in a complex with the laminin- and collagen-I-binding α3β1 integrin. We also find that JAM-A interacts with tetraspanins CD151 and CD9, which both interact with α3β1 integrin and regulate α3β1 integrin activity in different contexts. Mapping experiments indicate that JAM-A associates with α3β1 integrin and tetraspanins CD151 and CD9 through its extracellular domain. Similar to depletion of JAM-A, depletion of either α3β1 integrin or tetraspanins CD151 and CD9 in MDCK cells slows down collective cell migration. Our findings suggest that JAM-A exists with α3β1 integrin and tetraspanins CD151 and CD9 in a functional complex to regulate collective cell migration of polarized epithelial cells.

Loss of contact inhibition of locomotion in the absence of JAM-A promotes entotic cell engulfment

“…The formation of cell-in-cell structures after JAM-A depletion required Rho-associated kinase (ROCK) activity ( Figure 2 D), which has been described to be necessary for entotic cell engulfment ( Overholtzer et al., 2007 ). JAM-A KD – WT collisions resulted predominantly in cell-in-cell structures in which the WT cells were engulfed by the JAM-A KD cells ( Figure 2 E), suggesting that the increased motility of single cells combined with a loss of contact inhibition observed after JAM-A depletion ( Kummer et al., 2022 ; Tholmann et al., 2022 ) prones cells to be invaded by other cells. Similar to what has been observed in the regulation of CIL ( Kummer et al., 2022 ), we observed no difference in the rate of entosis between heterotypic JAM-A KD vs WT collisions and homotypic JAM-A KD vs JAM-A KD collisions ( Figure S2 ), further supporting that trans -homphilic JAM-A interaction is required during entosis.…”

Methamphetamine induces transcriptional changes in cultured HIV-infected mature monocytes that may contribute to HIV neuropathogenesis

“…CD151 also forms homotypic interactions that lead to increased cell movement and MMP-9 production ( 66 ). CD151 can interact with JAM-A, a molecule increased on the surface of HIV-infected mature monocytes, contributing to increased cell migration ( 33 , 67 ). Changes in these molecules may contribute to cell adhesion, downstream signaling activation, and their consequent migration from the periphery into the CNS.…”

Potential diagnostic of lymph node metastasis and prognostic values of TM4SFs in papillary thyroid carcinoma patients

“…The TM4SF protein family includes at least 16 members, most of which are leukocyte surface proteins, and the superfamily is characterized by four highly conserved transmembrane domains of cell surface proteins, making TM4SF proteins especially well adapted in tumor invasion and migration. The tetraspanins, CD151 and CD9, were previously found to interact with JAM-A and directly regulate α3β1 integrin activity to promote tumor invasion and migration ( Thölmann et al, 2022 ). Recent evidence suggested that tetraspanins have an important impact on mitochondria turnover and regulation of cellular metabolism, which can promote metastasis ( Toribio and Yáñez-Mó, 2022 ).…”