Complexes of plexin-A4 and plexin-D1 convey semaphorin-3C signals to induce cytoskeletal collapse in the absence of neuropilins

Class-3 semaphorin guidance factors bind to receptor complexes containing neuropilin and plexin receptors. A semaphorin may bind to several receptor complexes containing somewhat different constituents, resulting in diverse effects on cell migration. U87MG glioblastoma cells express both neuropilins and the four class-A plexins. They respond by cytoskeletal collapse and cell contraction to sema3A or sema3B but fail to contract in response to Sema3C, Sema3D, Sema3G or sema3E even when class-A plexins are over-expressed in the cells. In-contrast, expression of recombinant plexin-D1 enabled contraction in response to these semaphorins. Surprisingly, unlike sema3D and sema3G, sema3C also induced the contraction and repulsion of plexin-D1 expressing U87MG cells in which both neuropilins were knocked-out using CRISPR/cas9. In the absence of neuropilins the EC-50 of sema3C was 5.5 fold higher, indicating that the neuropilins function as enhancers of plexin-D1 mediated sema3C signaling but are not absolutely required for sema3C signal transduction. Interestingly, in the absence of neuropilins, plexin-A4 formed complexes with plexin-D1, and was required in addition to plexin-D1 to enable sema3C induced signal transduction.