Skip to main content
Fig. 6 | Cell Communication and Signaling

Fig. 6

From: Human hyaluronic acid synthase-1 promotes malignant transformation via epithelial-to-mesenchymal transition, micronucleation and centrosome abnormalities

Fig. 6

HAS1-synthesized HA interacts with RHAMM. a RHAMM and its splice variants are associated with cellular HA (synthesized from HAS1 overexpression) during mitosis and G1/S phase. HeLa cells were co-transfected with plasmids expressing A2-tagged HAS1 (A2-HAS1) and full-length GFP-tagged RHAMM (RHAMM-GFP) or the splice-variants of RHAMM (RHAMM-Ex4-GFP). Cell populations were synchronized in Mitosis or G1/S using thymidine block and synchronization was verified using flow cytometry (Supplementary Fig. 3B). Total cellular HA was isolated using biotinylated bovine HA-binding-protein and streptavidin-conjugated magnetic beads. The isolated beads were treated (+) with hyaluronidase (HAase). Samples were subjected to immunoblotting for RHAMM and HAS1. b BRCA1 interacted with RHAMM isoforms but not with the other HA-binding protein Neurocan. GFP-tagged RHAMM isoforms and GFP-ΔNeurocan were expressed in HeLa cells and HA-binding proteins were isolated from the cell lysates using biotinylated-HA as “bait”. Immunoblotting of pull-down material with and without HAase treatment revealed that endogenous BRCA1 were found to be associated with RHAMM isoforms (but not with Neurocan), suggesting that BRCA1 may interact directly with RHAMM

Back to article page