Ubiquitin-like protein MNSFβ noncovalently binds to molecular chaperone HSPA8 and regulates osteoclastogenesis

Abstract

MNSFβ, a ubiquitin-like protein, covalently binds to various target proteins including proapoptotic Bcl-G. During the course of isolation of MNSFβ-conjugating enzyme(s), we identified a novel target protein for MNSFβ. MALDI-TOF MS fingerprinting revealed that the MNSFβ-interacting protein is HSPA8 (heat shock 70-kDa protein 8). We observed that MNSFβ noncovalently binds to HSPA8 in the presence of ATP in vitro. Double knockdown of MNSFβ and HSPA8 strongly inhibited RANKL-induced osteoclastogenesis from Raw264.7 macrophage-like cells. The same treatment inhibited RANKL-induced ERK1/2 and p38 phosphorylation and TNFα production, suggesting that the association of MNSFβ with HSPA8 may promote RANKL-induced osteoclastogenesis. This is the first report that MNSFβ binds to a protein substrate via the noncovalent association and exerts biological effects.

Communications Between Bone Marrow Macrophages and Bone Cells in Bone Remodeling

“…NF-kB, composed of subunits such as p65, is a downstream transcription factor of the RANKL-RANKL signaling pathway (Park et al, Interfere the PTEN/PI3K/AKT signaling pathway to regulate the biological behavior of cells Zheng et al, 2017;Yang et al, 2018 mir-155 Promote the secretion of inflammatory cytokines Stanczyk et al, 2008;Heymans et al, 2013;Ying et al, 2017 mir-5106 Inhibit the genes SIK2 and SIK3 to promote bone formation Xiong et al, 2020 Alarmins Annexins Induce bone resorption and help macrophages to work Li et al, 2005;D'Souza et al, 2012;Stukes et al, 2016;McArthur et al, 2020;Xia et al, 2020;Xiong et al, 2020 Galectins Act on the differentiation of various bone cells Andersen et al, 2003;Shimura et al, 2005;Nakajima et al, 2014;Weilner et al, 2016;Simon et al, 2017;Iacobini et al, 2018 HSP; fibronectin Functions through osteoblasts and osteoclasts. Moursi et al, 1996;Koh et al, 2009;Gramoun et al, 2010;Notsu et al, 2016;Nakamura et al, 2019 2017). Nuclear factor of activated T cells cytoplasmic 1 (NFATc1) is a master regulator of osteoclast differentiation (Okamoto and Takayanagi, 2011).…”

Macrophage-Derived Extracellular Vesicles as Carriers of Alarmins and Their Potential Involvement in Bone Homeostasis

“…In contrast, the HSP70 family member heat-shock 70-kDA protein-8 binds to the ubiquitin-like protein monoclonal non-specific suppressor factor β and double knockdown of these factors inhibited RANKL-induced osteoclastogenesis (64). The effects of HSP90 on osteoclastogenesis are more controversial.…”

Subchronic intravenous toxicity study of biofunctional ZnO and its application as a fluorescence probe for cell‐specific targeting

“…We synthesized ZHIE NPs‐labeled anti‐Mac1 Ab (ZHIE‐Mac1Ab) (Figure ) and examined their staining ability for Mac1‐expressing Raw264.7 cells, which have been studied in our laboratory . The fluorescence images were taken after 1‐hour incubation of Raw264.7 cells with ZHIE‐Mac1Ab solution (10 nM).…”