Functional characterization of lysophosphatidic acid receptor 1 mutants identified in rat cancer tissues

Lysophosphatidic acid (LPA), an extracellular lipid mediator, exerts various cellular effects through activation of LPA receptors, LPA₁-LPA₆, in many types of cells including cancer cells. We recently found several missense mutations of Lpar1 in rat cancer tissues. One of these mutations is located at the extracellular tip of the seventh transmembrane domain of LPA₁, and another three mutations are found within the NPXXY motif in the seventh transmembrane domain. These mutants are designated F295S LPA₁ and P308S, I310T, and Y311H LPA₁, respectively. Here, we examined the functions of these LPA₁ mutants. Compared with wild-type (WT) LPA₁, F295S, P308S, and I310T LPA₁ showed decreased maximal responses in inhibition of cAMP formation, Ca²⁺ mobilization, and cytoskeletal changes. Y311H LPA₁ failed to show LPA-induced cellular responses. However, these LPA₁ mutants were internalized in response to LPA exposure. Finally, while WT and F295S LPA₁ showed a similar, broad distribution throughout the cell, P308S, I310T, and Y311H LPA₁ displayed a restricted cellular distribution and co-localized with the endoplasmic reticulum. These data suggest that the LPA₁ mutants perturb LPA signaling in cancer tissues.