Breast cancer-associated gene 3 interacts with Rac1 and augments NF-κB signaling in vitro, but has no effect on RANKL-induced bone resorption in vivo

Yao,Chen; Yu,Kuan-Ping; Philbrick,William; Sun,Ben-Hua; Simpson,Christine; Zhang,Changqing; Insogna,Karl

doi:10.3892/ijmm.2017.3091

Breast cancer-associated gene 3 interacts with Rac1 and augments NF-κB signaling in vitro, but has no effect on RANKL-induced bone resorption in vivo

Authors:
- Chen Yao
- Kuan-Ping Yu
- William Philbrick
- Ben-Hua Sun
- Christine Simpson
- Changqing Zhang
- Karl Insogna
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Affiliations: Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USA, Department of Orthopaedics, Shanghai Jiaotong University Affiliated Shanghai No. 6 People's Hospital, Shanghai 200233, P.R. China
Published online on: August 4, 2017 https://doi.org/10.3892/ijmm.2017.3091
Pages: 1067-1077
Copyright: © Yao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Breast cancer-associated gene 3 (BCA3) is a recently identified adaptor protein whose functions are still being defined. BCA3 has been reported to be an important regulator of nuclear factor-κB (NF-κB) signaling. It has also been reported to interact with the small GTPase, Rac1. Consistent with that observation, in the present study, BCA3 was found to interact with nuclear Rac1 in 293 cells and influence NF-κB signaling. Additional experiments revealed that depending on cell type, BCA3 augmented, attenuated or had no effect on NF-κB signaling in vitro. Since canonical NF-κB signaling is a critical downstream target from activated receptor activator of nuclear factor κB (RANK) that is required for mature osteoclast formation and function, BCA3 was selectively overexpressed in osteoclasts in vivo using the cathepsin K promoter and the response to exogenous receptor activator of nuclear factor κB ligand (RANKL) administration was examined. Despite its ability to augment NF-κB signaling in other cells, transgenic animals injected with high-dose RANKL had the same hypercalcemic response as their wild‑type littermates. Furthermore, the degree of bone loss induced by a 2-week infusion of low-dose RANKL was the same in both groups. Combined with earlier studies, the data from our study data indicate that BCA3 can affect NF-κB signaling and that BCA3 plays a cell-type dependent role in this process. The significance of the BCA3/NF-κB interaction in vivo in bone remains to be determined.

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1	Kitching R, Li H, Wong MJ, Kanaganayakam S, Kahn H and Seth A: Characterization of a novel human breast cancer associated gene (BCA3) encoding an alternatively spliced proline-rich protein. Biochim Biophys Acta. 1625:116–121. 2003. View Article : Google Scholar : PubMed/NCBI
2	Sastri M, Barraclough DM, Carmichael PT and Taylor SS: A-kinase-interacting protein localizes protein kinase A in the nucleus. Proc Natl Acad Sci USA. 102:349–354. 2005. View Article : Google Scholar : PubMed/NCBI
3	Gao N, Asamitsu K, Hibi Y, Ueno T and Okamoto T: AKIP1 enhances NF-kappaB-dependent gene expression by promoting the nuclear retention and phosphorylation of p65. J Biol Chem. 283:7834–7843. 2008. View Article : Google Scholar : PubMed/NCBI
4	Gao N, Hibi Y, Cueno M, Asamitsu K and Okamoto T: A-kinase-interacting protein 1 (AKIP1) acts as a molecular determinant of PKA in NF-kappaB signaling. J Biol Chem. 285:28097–28104. 2010. View Article : Google Scholar : PubMed/NCBI
5	King CC, Sastri M, Chang P, Pennypacker J and Taylor SS: The rate of NF-κB nuclear translocation is regulated by PKA and A kinase interacting protein 1. PLoS One. 6:e187132011. View Article : Google Scholar
6	Leon DA and Cànaves JM: In silico study of breast cancer associated gene 3 using LION Target Engine and other tools. Biotechniques. 35:1222–1226. 12281230–1231. 2003.PubMed/NCBI
7	Leung TH and Ngan HY: Interaction of TAp73 and breast cancer-associated gene 3 enhances the sensitivity of cervical cancer cells in response to irradiation-induced apoptosis. Cancer Res. 70:6486–6496. 2010. View Article : Google Scholar : PubMed/NCBI
8	Qin HY and Han H: Effect of KyoT2 binding protein KBP1 on RBP-J-mediated transcriptional activity. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 20:544–547. 2004.In Chinese. PubMed/NCBI
9	Gao F, Cheng J, Shi T and Yeh ET: Neddylation of a breast cancer-associated protein recruits a class III histone deacetylase that represses NFkappaB-dependent transcription. Nat Cell Biol. 8:1171–1177. 2006. View Article : Google Scholar : PubMed/NCBI
10	Sastri M, Haushalter KJ, Panneerselvam M, Chang P, Fridolfsson H, Finley JC, Ng D, Schilling JM, Miyanohara A, Day ME, et al: A kinase interacting protein (AKIP1) is a key regulator of cardiac stress. Proc Natl Acad Sci USA. 110:E387–E396. 2013. View Article : Google Scholar : PubMed/NCBI
11	Hashimoto M, Murata E and Aoki T: Secretory protein with RING finger domain (SPRING) specific to Trypanosoma cruzi is directed, as a ubiquitin ligase related protein, to the nucleus of host cells. Cell Microbiol. 12:19–30. 2010. View Article : Google Scholar
12	Yu KP, Itokawa T, Zhu ML, Syam S, Seth A and Insogna K: Breast cancer-associated gene 3 (BCA3) is a novel Rac1-interacting protein. J Bone Miner Res. 22:628–637. 2007. View Article : Google Scholar : PubMed/NCBI
13	Sakai H, Chen Y, Itokawa T, Yu KP, Zhu ML and Insogna K: Activated c-Fms recruits Vav and Rac during CSF-1-induced cytoskeletal remodeling and spreading in osteoclasts. Bone. 39:1290–1301. 2006. View Article : Google Scholar : PubMed/NCBI
14	Diebold I, Djordjevic T, Hess J and Görlach A: Rac-1 promotes pulmonary artery smooth muscle cell proliferation by upregulation of plasminogen activator inhibitor-1: role of NFkappaB-dependent hypoxia-inducible factor-1alpha transcription. Thromb Haemost. 100:1021–1028. 2008.
15	Williams LM, Lali F, Willetts K, Balague C, Godessart N, Brennan F, Feldmann M and Foxwell BM: Rac mediates TNF-induced cytokine production via modulation of NF-kappaB. Mol Immunol. 45:2446–2454. 2008. View Article : Google Scholar : PubMed/NCBI
16	Yu M, Qi X, Moreno JL, Farber DL and Keegan AD: NF-κB signaling participates in both RANKL- and IL-4-induced macrophage fusion: Receptor cross-talk leads to alterations in NF-κB pathways. J Immunol. 187:1797–1806. 2011. View Article : Google Scholar : PubMed/NCBI
17	Yamashita T, Yao Z, Li F, Zhang Q, Badell IR, Schwarz EM, Takeshita S, Wagner EF, Noda M, Matsuo K, et al: NF-kappaB p50 and p52 regulate receptor activator of NF-kappaB ligand (RANKL) and tumor necrosis factor-induced osteoclast precursor differentiation by activating c-Fos and NFATc1. J Biol Chem. 282:18245–18253. 2007. View Article : Google Scholar : PubMed/NCBI
18	Valenzuela DM, Murphy AJ, Frendewey D, Gale NW, Economides AN, Auerbach W, Poueymirou WT, Adams NC, Rojas J, Yasenchak J, et al: High-throughput engineering of the mouse genome coupled with high-resolution expression analysis. Nat Biotechnol. 21:652–659. 2003. View Article : Google Scholar : PubMed/NCBI
19	Copeland NG, Jenkins NA and Court DL: Recombineering: A powerful new tool for mouse functional genomics. Nat Rev Genet. 2:769–779. 2001. View Article : Google Scholar : PubMed/NCBI
20	Liu P, Jenkins NA and Copeland NG: A highly efficient recombineering-based method for generating conditional knockout mutations. Genome Res. 13:476–484. 2003. View Article : Google Scholar : PubMed/NCBI
21	Sarov M, Schneider S, Pozniakovski A, Roguev A, Ernst S, Zhang Y, Hyman AA and Stewart AF: A recombineering pipeline for functional genomics applied to Caenorhabditis elegans. Nat Methods. 3:839–844. 2006. View Article : Google Scholar : PubMed/NCBI
22	Rohrschneider LR, Rothwell VM and Nicola NA: Transformation of murine fibroblasts by a retrovirus encoding the murine c-fms proto-oncogene. Oncogene. 4:1015–1022. 1989.PubMed/NCBI
23	Lacey DL, Timms E, Tan HL, Kelley MJ, Dunstan CR, Burgess T, Elliott R, Colombero A, Elliott G, Scully S, et al: Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell. 93:165–176. 1998. View Article : Google Scholar : PubMed/NCBI
24	Yao GQ, Wu JJ, Troiano N, Zhu ML, Xiao XY and Insogna K: Selective deletion of the membrane-bound colony stimulating factor 1 isoform leads to high bone mass but does not protect against estrogen-deficiency bone loss. J Bone Miner Metab. 30:408–418. 2012. View Article : Google Scholar
25	Kawano T, Zhu M, Troiano N, Horowitz M, Bian J, Gundberg C, Kolodziejczak K and Insogna K: LIM kinase 1 deficient mice have reduced bone mass. Bone. 52:70–82. 2013. View Article : Google Scholar :
26	Perona R, Montaner S, Saniger L, Sánchez-Pérez I, Bravo R and Lacal JC: Activation of the nuclear factor-kappaB by Rho, CDC42, and Rac-1 proteins. Genes Dev. 11:463–475. 1997. View Article : Google Scholar : PubMed/NCBI
27	Montaner S, Perona R, Saniger L and Lacal JC: Multiple signalling pathways lead to the activation of the nuclear factor kappaB by the Rho family of GTPases. J Biol Chem. 273:12779–12785. 1998. View Article : Google Scholar : PubMed/NCBI
28	Miyazaki T, Katagiri H, Kanegae Y, Takayanagi H, Sawada Y, Yamamoto A, Pando MP, Asano T, Verma IM, Oda H, et al: Reciprocal role of ERK and NF-kappaB pathways in survival and activation of osteoclasts. J Cell Biol. 148:333–342. 2000. View Article : Google Scholar : PubMed/NCBI
29	Ogasawara T1, Katagiri M, Yamamoto A, Hoshi K, Takato T, Nakamura K, Tanaka S, Okayama H and Kawaguchi H: Osteoclast differentiation by RANKL requires NF-kappaB-mediated down-regulation of cyclin-dependent kinase 6 (Cdk6). J Bone Miner Res. 19:1128–1136. 2004. View Article : Google Scholar : PubMed/NCBI
30	Yao C, Yao GQ, Sun BH, Zhang C, Tommasini SM and Insogna K: The transcription factor T-box3 regulates colony stimulating factor 1-dependent Jun dimerization protein 2 expression and plays an important role in osteoclastogenesis. J Biol Chem. 289:6775–6790. 2014. View Article : Google Scholar : PubMed/NCBI
31	Lloyd SA, Yuan YY, Kostenuik PJ, Ominsky MS, Lau AG, Morony S, Stolina M, Asuncion FJ and Bateman TA: Soluble RANKL induces high bone turnover and decreases bone volume, density, and strength in mice. Calcif Tissue Int. 82:361–372. 2008. View Article : Google Scholar : PubMed/NCBI
32	Frost JA, Swantek JL, Stippec S, Yin MJ, Gaynor R and Cobb MH: Stimulation of NFkappa B activity by multiple signaling pathways requires PAK1. J Biol Chem. 275:19693–19699. 2000. View Article : Google Scholar : PubMed/NCBI
33	Boyer L, Travaglione S, Falzano L, Gauthier NC, Popoff MR, Lemichez E, Fiorentini C and Fabbri A: Rac GTPase instructs nuclear factor-kappaB activation by conveying the SCF complex and IkBalpha to the ruffling membranes. Mol Biol Cell. 15:1124–1133. 2004. View Article : Google Scholar :
34	Michaelson D, Abidi W, Guardavaccaro D, Zhou M, Ahearn I, Pagano M and Philips MR: Rac1 accumulates in the nucleus during the G2 phase of the cell cycle and promotes cell division. J Cell Biol. 181:485–496. 2008. View Article : Google Scholar : PubMed/NCBI
35	van Hennik PB, ten Klooster JP, Halstead JR, Voermans C, Anthony EC, Divecha N and Hordijk PL: The C-terminal domain of Rac1 contains two motifs that control targeting and signaling specificity. J Biol Chem. 278:39166–39175. 2003. View Article : Google Scholar : PubMed/NCBI
36	Sandrock K, Bielek H, Schradi K, Schmidt G and Klugbauer N: The nuclear import of the small GTPase Rac1 is mediated by the direct interaction with karyopherin alpha2. Traffic. 11:198–209. 2010. View Article : Google Scholar
37	Vaira S, Alhawagri M, Anwisye I, Kitaura H, Faccio R and Novack DV: RelA/p65 promotes osteoclast differentiation by blocking a RANKL-induced apoptotic JNK pathway in mice. J Clin Invest. 118:2088–2097. 2008.PubMed/NCBI
38	Ikeda F, Matsubara T, Tsurukai T, Hata K, Nishimura R and Yoneda T: JNK/c-Jun signaling mediates an anti-apoptotic effect of RANKL in osteoclasts. J Bone Miner Res. 23:907–914. 2008. View Article : Google Scholar : PubMed/NCBI