NOD1 cooperates with HAX‐1 to promote cell migration in a RIPK2‐ and NF‐ĸB‐independent manner

Abstract

The human Nod‐like receptor protein NOD1 is a well‐described pattern‐recognition receptor (PRR) with diverse functions. NOD1 associates with F‐actin and its protein levels are upregulated in metastatic cancer cells. A hallmark of cancer cells is their ability to migrate, which involves actin remodelling. Using chemotaxis and wound healing assays, we show that NOD1 expression correlated with the migration rate and chemotactic index in the cervical carcinoma cell line HeLa. The effect of NOD1 in cell migration was independent of the downstream kinase RIPK2 and NF‐ĸB activity. Additionally, NOD1 negatively regulated the phosphorylation status of cofilin, which inhibits actin turnover. Co‐immunoprecipitation assays identified HCLS1‐associated protein X‐1 (HAX‐1) as a previously unknown interaction partner of NOD1. Silencing of HAX‐1 expression reduced the migration behaviour to similar levels as NOD1 knockdown, and simultaneous knockdown of NOD1 and HAX‐1 showed no additive effect, suggesting that both proteins act in the same pathway. In conclusion, our data revealed an important role of the PRR NOD1 in regulating cell migration as well as chemotaxis in human cervical cancer cells and identified HAX‐1 as a protein that interacts with NOD1 and is involved in this signalling pathway.

Label-free detection of prostaglandin transporter (SLCO2A1) function and inhibition: insights by wound healing and TRACT assays

“…Thereby this method provides a more quantitative method to evaluate wound closure, which is followed in real time. Hezinger et al (2023) provided evidence that impedance-based scratch assays yield similar results as the traditional microscopy-based wound healing assay as observed for NOD1 silencing in HeLa cells ( Hezinger et al, 2023 ). Wound healing occurs more rapidly when extracellular PGE 2 levels are increased either due to direct addition of 100 nM PGE 2 or by a decrease in PGT mediated uptake and breakdown of PGE 2 due to T26A mediated inhibition of PGT ( Figure 5 ).…”