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Cloning, sequencing, and functional characterization of the rat homologue of receptor activator of NF-kappaB ligand.

A complementary DNA (cDNA) encoding the rat homologue of receptor activator of NF-kappaB ligand/ osteoprotegerin ligand/ osteoclast differentiation factor/tumor necrosis factor (TNF)-related activation-induced cytokine ( RANKL/ OPGL/ ODF/TRANCE) was cloned and sequenced from tibias of ovariectomized (OVX) rats. The predicted amino acid sequence of rat RANKL (rRANKL) has 84% and 96% identity to that of human and mouse RANKL, respectively, and 35% and 37% similarity to that of human and mouse TNF-related apoptosis-inducing ligand (TRAIL), respectively. RANKL transcripts were expressed abundantly in the thymus and bone tissues of OVX rats. rRANKL has a single hydrophobic region between residues 53 and 69, which is most likely to serve as a transmembrane domain. The long C-terminal region containing beta-sheet-forming sequences of the TNF-like core is considered the extracellular region. Three truncated domains within the TNF-like core region were expressed as glutathione S-transferase ( GST) fusion proteins and investigated for their ability to induce osteoclastogenesis. The results showed that GST-rRANKL (aa160-318) containing the full TNF-like core region had the highest capability to induce the formation of osteoclast-like cells from RAW264.7 cells. GST-rRANKL (aa239-318 and aa160-268) had lesser degrees of osteoclast inductivity. Furthermore, the GST-rRANKL (aa160-318) is capable of (1) inducing osteoclast formation from rat spleen cells in the presence of macrophage colony-stimulating factor ( M-CSF), (2) stimulating mature rat osteoclast polarization and bone resorption ex vivo, and (3) inducing systemic hypercalcemia in vivo; thus the full TNF-like core region of rRANKL is an important regulator of calcium homeostasis and osteoclastic function.[1]

References

  1. Cloning, sequencing, and functional characterization of the rat homologue of receptor activator of NF-kappaB ligand. Xu, J., Tan, J.W., Huang, L., Gao, X.H., Laird, R., Liu, D., Wysocki, S., Zheng, M.H. J. Bone Miner. Res. (2000) [Pubmed]
 
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