We believe that the dynamic exchange of subunits plays a role in the regulation of ULF assembly and the maintenance of a soluble vimentin pool during the reorganization of filament networks. After all pcFPs have been read out, the still-intact paFP. Furthermore, vimentin exchange in ULFs required ATP, and ATP depletion caused a dramatic reduction of the soluble tetramer pool. mEos3.2-A69T 14, is imaged using a combination of 488 and 730 nm light for photoconversion. Our data demonstrate that cells expressing vimentinY117L contained a large pool of soluble vimentin tetramers that was in rapid equilibrium with ULFs. Subunit exchange among ULFs occurred within seconds and was limited by the diffusion of soluble subunits in the cytoplasm rather than by the association and dissociation of subunits from ULFs. By tagging vimentinY117L with a photoconvertible protein mEos3.2 and photoconverting ULFs in a limited area of the cytoplasm, we found that ULFs, unlike mature filaments, were highly dynamic. To block vimentin assembly at the ULF stage, we used the Y117L vimentin mutant (vimentinY117L). Here, we studied the dynamics of subunit exchange in VIF precursors, called unit-length filaments (ULFs), formed by the lateral association of eight vimentin tetramers. For example, recent studies suggest that mature vimentin IFs (VIFs) undergo rearrangement by severing and reannealing, but direct subunit exchange within the filament plays little role in filament dynamics at steady state. Various mechanisms were proposed to be responsible for this plasticity depending on the type of IF polymer and the biological context. Intermediate filaments (IFs) are a component of the cytoskeleton capable of profound reorganization in response to specific physiological situations, such as differentiation, cell division, and motility.
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