Conformational mechanism for the stability of microtubule-kinetochore attachments

Regulating the stability of microtubule(MT)-kinetochore attachments is fundamental to avoiding mitotic errors and ensure proper chromosome segregation during cell division. While biochemical factors involved in this process have been identified, its mechanics still needs to be better understood. Here we introduce and simulate a mechanical model of MT-kinetochore interactions in which the stability of the attachment is ruled by the geometrical conformations of curling MT-protofilaments entangled in kinetochore fibrils. The model allows us to reproduce with good accuracy in vitro experimental measurements of the detachment times of yeast kinetochores from MTs under external pulling forces. Numerical simulations suggest that geometrical features of MT-protofilaments may play an important role in the switch between stable and unstable attachments

Anaphase B.

Anaphase B spindle elongation is characterized by the sliding apart of overlapping antiparallel interpolar (ip) microtubules (MTs) as the two opposite spindle poles separate, pulling along disjoined sister chromatids, thereby contributing to chromosome segregation and the propagation of all cellular life. The major biochemical "modules" that cooperate to mediate pole-pole separation include: (i) midzone pushing or (ii) braking by MT crosslinkers, such as kinesin-5 motors, which facilitate or restrict the outward sliding of antiparallel interpolar MTs (ipMTs); (iii) cortical pulling by disassembling astral MTs (aMTs) and/or dynein motors that pull aMTs outwards; (iv) ipMT plus end dynamics, notably net polymerization; and (v) ipMT minus end depolymerization manifest as poleward flux. The differential combination of these modules in different cell types produces diversity in the anaphase B mechanism. Combinations of antagonist modules can create a force balance that maintains the dynamic pre-anaphase B spindle at constant length. Tipping such a force balance at anaphase B onset can initiate and control the rate of spindle elongation. The activities of the basic motor filament components of the anaphase B machinery are controlled by a network of non-motor MT-associated proteins (MAPs), for example the key MT cross-linker, Ase1p/PRC1, and various cell-cycle kinases, phosphatases, and proteases. This review focuses on the molecular mechanisms of anaphase B spindle elongation in eukaryotic cells and briefly mentions bacterial DNA segregation systems that operate by spindle elongation

Mitotic force generators and chromosome segregation

The mitotic spindle uses dynamic microtubules and mitotic motors to generate the pico-Newton scale forces that are needed to drive the mitotic movements that underlie chromosome capture, alignment and segregation. Here, we consider the biophysical and molecular basis of force-generation for chromosome movements in the spindle, and, with reference to the Drosophila embryo mitotic spindle, we briefly discuss how mathematical modeling can complement experimental analysis to illuminate the mechanisms of chromosome-to-pole motility during anaphase A and spindle elongation during anaphase B

Distribution of lifetimes of kinetochore-microtubule attachments: interplay of energy landscape, molecular motors and microtubule (de-)polymerization

Before a cell divides into two daughter cells, chromosomes are replicated resulting in two sister chromosomes embracing each other. Each sister chromosome is bound to a separate proteinous structure, called kinetochore (kt), that captures the tip of a filamentous protein, called microtubule (MT). Two oppositely oriented MTs pull the two kts attached to two sister chromosomes thereby pulling the two sisters away from each other. Here we theoretically study an even simpler system, namely an isolated kt coupled to a single MT; this system mimics an {\it in-vitro} experiment where a single kt-MT attachment is reconstituted using purified extracts from budding yeast. Our models not only account for the experimentally observed "catch-bond-like" behavior of the kt-MT coupling, but also make new predictions on the probability distribution of the lifetimes of the attachments. In principle, our new predictions can be tested by analyzing the data collected in the {\it in-vitro} experiments provided the experiment is repeated sufficiently large number of times. Our theory provides a deep insight into the effects of (a) size, (b) energetics, and (c) stochastic kinetics of the kt-MT coupling on the distribution of the lifetimes of these attachments.Comment: This is an author-created, un-copyedited version of an article accepted for publication in "Physical Biology" (IOP). IOP Publishing Ltd is not responsible for any errors or omissions in this version of the manuscript or any version derived from i

Doctor of Philosophy

dissertationCell division is a complex process that involves carefully orchestrated chemical and mechanical events. Tight regulation is vital during division, since a breakdown in control mechanisms can lead to serious disorders such as cancer. A key step in division is the movement of chromosomes to specific locations in the cell with remarkable precision. In higher eukaryotes, the movement of chromosomes has been well observed over the course of hundreds of years. Yet, the mechanisms underlying chromosome motility and the control of precise chromosome localizations in the cell are poorly understood. More recently, a wealth of experimental data has become available for bacterial division. Despite the long supported theory that bacteria and eukaryotes differ widely when undergoing division, it is emerging that similar mechanisms for motility and cell cycle control might be at play in both cell types. Mathematical modeling is useful in the study of these dynamic cellular environments, where it is difficult to experimentally uncover the mechanisms that drive a multitude of mechanical and chemical events. In this dissertation, we develop various mathematical models that address the question of how dynamic polymers can move large objects such as chromosomes in higher eukaryotes and in bacteria. Then, we develop models that address how chemical and mechanical signals can be coordinated to control the precise localization of a chromosome. The mathematical models proposed here employ stochastic differential equations, ordinary differential equations and partial differential equations. The models are numerically simulated to obtain solutions for various parameter values, but we also use tools from bifurcation theory, asymptotic and perturbation methods for our model analysis. Our mathematical models can not only reproduce the experimental data at hand, but also make predictions about the mechanisms underlying chromosome motility in dividing cells