Cloning, Sequencing, and Functional Characterization of the Rat Homologue of Receptor Activator of NF‐κB Ligand

A complementary DNA (cDNA) encoding the rat homologue of receptor activator of NF‐κB 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) ra...

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Veröffentlicht in:Journal of bone and mineral research 2000-11, Vol.15 (11), p.2178-2186
Hauptverfasser: Xu, Jiake, Tan, Jamie Wy, Huang, Lin, Gao, Xiu‐Hui, Laird, Rebecca, Liu, Dan, Wysocki, Stan, Zheng, Ming H.
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container_end_page 2186
container_issue 11
container_start_page 2178
container_title Journal of bone and mineral research
container_volume 15
creator Xu, Jiake
Tan, Jamie Wy
Huang, Lin
Gao, Xiu‐Hui
Laird, Rebecca
Liu, Dan
Wysocki, Stan
Zheng, Ming H.
description A complementary DNA (cDNA) encoding the rat homologue of receptor activator of NF‐κB 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 β‐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.
doi_str_mv 10.1359/jbmr.2000.15.11.2178
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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 β‐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.</description><identifier>ISSN: 0884-0431</identifier><identifier>EISSN: 1523-4681</identifier><identifier>DOI: 10.1359/jbmr.2000.15.11.2178</identifier><identifier>CODEN: JBMREJ</identifier><language>eng</language><publisher>Washington, DC: John Wiley and Sons and The American Society for Bone and Mineral Research (ASBMR)</publisher><subject>Biological and medical sciences ; bone resorption ; Fundamental and applied biological sciences. 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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.</description><subject>Biological and medical sciences</subject><subject>bone resorption</subject><subject>Fundamental and applied biological sciences. 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Psychology</topic><topic>hypercalcemia</topic><topic>Medical sciences</topic><topic>Metabolic diseases</topic><topic>Miscellaneous</topic><topic>osteoclast</topic><topic>osteoprotegerin</topic><topic>Other metabolic disorders</topic><topic>RANKL protein</topic><topic>receptor activator of NK‐κB/osteoprotegerin ligand/osteoclast differentiation factor/tumor necrosis factor‐related activation‐induced cytokine</topic><topic>Skeleton and joints</topic><topic>TRANCE protein</topic><topic>Vertebrates: osteoarticular system, musculoskeletal system</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xu, Jiake</creatorcontrib><creatorcontrib>Tan, Jamie Wy</creatorcontrib><creatorcontrib>Huang, Lin</creatorcontrib><creatorcontrib>Gao, Xiu‐Hui</creatorcontrib><creatorcontrib>Laird, Rebecca</creatorcontrib><creatorcontrib>Liu, Dan</creatorcontrib><creatorcontrib>Wysocki, Stan</creatorcontrib><creatorcontrib>Zheng, Ming H.</creatorcontrib><collection>Pascal-Francis</collection><collection>CrossRef</collection><collection>Calcium &amp; Calcified Tissue Abstracts</collection><jtitle>Journal of bone and mineral research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xu, Jiake</au><au>Tan, Jamie Wy</au><au>Huang, Lin</au><au>Gao, Xiu‐Hui</au><au>Laird, Rebecca</au><au>Liu, Dan</au><au>Wysocki, Stan</au><au>Zheng, Ming H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning, Sequencing, and Functional Characterization of the Rat Homologue of Receptor Activator of NF‐κB Ligand</atitle><jtitle>Journal of bone and mineral research</jtitle><date>2000-11</date><risdate>2000</risdate><volume>15</volume><issue>11</issue><spage>2178</spage><epage>2186</epage><pages>2178-2186</pages><issn>0884-0431</issn><eissn>1523-4681</eissn><coden>JBMREJ</coden><abstract>A complementary DNA (cDNA) encoding the rat homologue of receptor activator of NF‐κB 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 β‐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.</abstract><cop>Washington, DC</cop><pub>John Wiley and Sons and The American Society for Bone and Mineral Research (ASBMR)</pub><doi>10.1359/jbmr.2000.15.11.2178</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record>
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source Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Access via Wiley Online Library; Oxford University Press Journals All Titles (1996-Current)
subjects Biological and medical sciences
bone resorption
Fundamental and applied biological sciences. Psychology
hypercalcemia
Medical sciences
Metabolic diseases
Miscellaneous
osteoclast
osteoprotegerin
Other metabolic disorders
RANKL protein
receptor activator of NK‐κB/osteoprotegerin ligand/osteoclast differentiation factor/tumor necrosis factor‐related activation‐induced cytokine
Skeleton and joints
TRANCE protein
Vertebrates: osteoarticular system, musculoskeletal system
title Cloning, Sequencing, and Functional Characterization of the Rat Homologue of Receptor Activator of NF‐κB Ligand
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