Viscoelastic Relaxation of the Nuclear Envelope Does Not Cause the Collapse of the Spindle After Ablation in S. pombe.

Abstract

A large molecular machine called the mitotic spindle is responsible for accurate chromosome segregation in eukaryotic cells. The spindle consists of protein filaments known as microtubules and microtubule-associated proteins such as motors and crosslinkers, which help impart its organization. In the case of the fission yeast S. pombe, these form a single bundle inside the nucleus. During spindle elongation, sliding by motor proteins provides an internal source of extensile forces, which are resisted by the compressive forces of the nuclear envelope. To probe the sources of this force balance, we cut the spindle using focused laser light at various stages of spindle elongation. We find that the spindle pole bodies collapse toward each other post-ablation. While this basic behavior has been previously observed, many questions remain about the timing, mechanics, and molecular requirements of this phenomenon. Here, we quantify the time scale of the relaxation and probe its underlying mechanism. We demonstrate that viscoelastic relaxation of the nuclear envelope cannot explain this phenomenon and provide evidence of active forces as the underlying mechanism.