{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T16:22:42Z","timestamp":1762100562191},"reference-count":56,"publisher":"Oxford University Press (OUP)","issue":"11","license":[{"start":{"date-parts":[[2006,11,1]],"date-time":"2006-11-01T00:00:00Z","timestamp":1162339200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2006,11,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>The Tcirg1 gene encodes the osteoclast-specific a3 isoform of the V-ATPase a subunit. Using the mouse osteoclastic model RAW264.7 cells, we studied Tcirg1 gene expression, and we identified PARP-1 as a transcriptional repressor negatively regulated by RANKL during osteoclastogenesis.<\/jats:p>\n               <jats:p>Introduction: The TCIRG1 gene encodes the a3 isoform of the V-ATPase a subunit, and mutations at this locus account for \u223c60% of infantile malignant osteopetrosis cases. Using RAW264.7 cells as an osteoclastic differentiation model, we undertook a transcriptional study of the mouse Tcirg1 gene focused on the 4-kb region upstream of the transcription starting point.<\/jats:p>\n               <jats:p>Materials and Methods: The promoter activity of serial-deletion fragments of the Tcirg1 gene promoter was monitored throughout the RAW264.7 cell differentiation process. We next performed EMSA, UV cross-linking, affinity purification, mass spectrometry analysis, gel supershift, and siRNA transfection experiments to identify the factor(s) interacting with the promoter.<\/jats:p>\n               <jats:p>Results: The \u22123946\/+113 region of the mouse Tcirg1 gene displayed a high basal promoter activity, which was enhanced by RANKL treatment of RAW264.7 cells. Constructs deleted up to \u22121589 retained this response to RANKL. A deletion up to \u22121402 induced a 3-fold enhancement of the basal activity, whereas RANKL response was not affected. EMSA experiments led us to identify within the \u22121589\/\u22121402 region, a 10-nucleotide sequence, which bound a nuclear protein present in nondifferentiated RAW264.7 cells. This interaction was lost using nuclear extracts derived from RANKL-treated cells. Affinity purification followed by mass spectrometry analysis and gel supershift assay allowed the identification of poly(ADP-ribose) polymerase-1 (PARP-1) as this transcriptional repressor, whereas Western blot experiments revealed the cleavage of the DNA-binding domain of PARP-1 on RANKL treatment. Finally, both PARP-1 depletion after siRNA transfection and RAW264.7 cell treatment by an inhibitor of PARP-1 activity induced an increase of a3 mRNA expression.<\/jats:p>\n               <jats:p>Conclusions: We provide evidence that the basal transcription activity of the Tcirg1 gene is negatively regulated by the binding of PARP-1 protein to its promoter region in mouse pre-osteoclast. On RANKL treatment, PARP-1 protein is cleaved and loses its repression effect, allowing an increase of Tcirg1 gene expression that is critical for osteoclast function.<\/jats:p>","DOI":"10.1359\/jbmr.060809","type":"journal-article","created":{"date-parts":[[2006,9,29]],"date-time":"2006-09-29T14:58:12Z","timestamp":1159541892000},"page":"1757-1769","source":"Crossref","is-referenced-by-count":23,"title":["RANKL Treatment Releases the Negative Regulation of the Poly(ADP-Ribose) Polymerase-1 on <i>Tcirg1<\/i> Gene Expression During Osteoclastogenesis"],"prefix":"10.1093","volume":"21","author":[{"given":"Guillaume E","family":"Beranger","sequence":"first","affiliation":[{"name":"G\u00e9PITOS\u2013K2943 CNRS\/UNSA, Facult\u00e9 de M\u00e9decine de l'Universit\u00e9 de Nice-Sophia Antipolis, Nice, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Momier","sequence":"additional","affiliation":[{"name":"G\u00e9PITOS\u2013K2943 CNRS\/UNSA, Facult\u00e9 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