A dynein independent role of Tctex-1 at the kinetochore

Abstract

Dynein light chains are accessory subunits of the cytoplasmic dynein complex, a minus-end directed microtubule motor. Here, we demonstrate that the dynein light chain Tctex-1 associates with unattached kinetochores and is essential for accurate chromosome segregation. Tctex-1 knockdown in cells does not affect the localization and function of dynein at the kinetochore, but produces a prolonged mitotic arrest with a few misaligned chromosomes, which are subsequently missegregated during anaphase. This function is independent of Tctex-1's association with dynein. The kinetochore localization of Tctex-1 is independent of the ZW10-dynein pathway, but requires the Ndc80 complex. Thus, our findings reveal a dynein independent role of Tctex-1 at the kinetochore to enhance the stability of kinetochore-microtubule attachment.

Non-Overlapping Distributions and Functions of the VDAC Family in Ciliogenesis

“…The potential role of VDAC1 on the spindle is beyond the scope of this study. However, it may be important to note that VDAC1 interacts with Tctex-1 [ 48 , 49 ], a light chain subunit of cytoplasmic dynein that also localizes to spindle poles, minus ends of spindle microtubules, and kinetochores during mitosis [ 50 ] and negatively regulates ciliogenesis [ 51 ]. In addition, VDAC1 located at the OMM and ER were able to bind to free tubulin [ 52 ], microtubules and MAP4, a microtubule associated protein that stabilizes microtubules [ 53 ].…”

Centrifugal Displacement of Nuclei Reveals Multiple LINC Complex Mechanisms for Homeostatic Nuclear Positioning

“…Cells were lysed in KLB buffer (25 mM Tris pH 7.4; 10 % glycerol; 150 mM NaCl;1% Triton X-100; 1 mM NaF; 1 mM Na 3 VO 4 ) with Halt™ protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific) and GFP-tagged proteins were immunoprecipitated with rabbit anti-GFP (Thermo Fisher Scientific) and Dynabeads®-protein G (Thermo Fisher Scientific) following a published method [67] with a minor change (incubation at 4 °C for 1 hr). Beads were washed with RIPA buffer (50 mM Tris HCl pH 8.0; 150 mM NaCl; 1.0% NP-40; 0.5% sodium deoxycholate; 0.1% SDS) and then bound proteins eluted with SDS sample buffer and analyzed by western blotting.…”

Diaphanous formin mDia2 regulates CENP-A levels at centromeres

“…Immunoblotting was performed as previously described ( Liu et al, 2015 ). In brief, cells were lysed in RIPA buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% NP-40, 0.1% SDS, and 0.5% Na-deoxycholate acid) and then denatured using SDS sample buffer.…”