Day, Richard N.Day, Kathleen H.Pavalko, Fredrick M.2023-04-172023-04-172021-12-23Day RN, Day KH, Pavalko FM. Direct visualization by FRET-FLIM of a putative mechanosome complex involving Src, Pyk2 and MBD2 in living MLO-Y4 cells. PLoS One. 2021;16(12):e0261660. Published 2021 Dec 23. doi:10.1371/journal.pone.0261660https://hdl.handle.net/1805/32429Earlier, we proposed the "mechanosome" concept as a testable model for understanding how mechanical stimuli detected by cell surface adhesion molecules are transmitted to modulate gene expression inside cells. Here, for the first time we document a putative mechanosome involving Src, Pyk2 and MBD2 in MLO-Y4 osteocytes with high spatial resolution using FRET-FLIM. Src-Pyk2 complexes were concentrated at the periphery of focal adhesions and the peri-nuclear region. Pyk2-MBD2 complexes were located primarily in the nucleus and peri-nuclear region. Lifetime measurements indicated that Src and MBD2 did not interact directly. Finally, mechanical stimulation by fluid flow induced apparent accumulation of Src-Pyk2 protein complexes in the peri-nuclear/nuclear region, consistent with the proposed behavior of a mechanosome in response to a mechanical stimulus.en-USAttribution 4.0 InternationalDNA-binding proteinsFluorescence resonance energy transferFocal adhesion kinase 2OsteocytesArticle