Anillin/Mid1p interacts with the ECSRT-associated protein Vps4p and mitotic kinases to regulate cytokinesis in fission yeast

Cytokinesis is the final stage of the cell cycle which separates cellular constituents to produce two daughter cells. Using Schizosaccharomyces pombe we have investigated the role of various classes of proteins involved in this process. Central to these is anillin/Mid1p which forms a ring-like structure at the cell equator that predicts the site of cell separation through septation in fission yeast. Here we demonstrate a direct physical interaction between Mid1p and the endosomal sorting complex required for transport (ESCRT)-associated protein Vps4p. The interaction is essential for cell viability, and Vps4p is required for the correct cellular localization of Mid1p. Furthermore, we show that Mid1p is phosphorylated by the aurora kinase Aurora A, that the interaction of mid1 and ark1 genes is essential for cell viability, and that Ark1p is also required for the correct cellular localization of Mid1p. We mapped the sites of phosphorylation of Mid1p by Aurora A and the polo kinase Plk1 and assessed their importance by mutational analysis. Mutational analysis revealed S332, S523 and S531 to be required for Mid1p function and its interaction with Vps4p, Ark1p and Plo1p. Combined our data suggest a physical interaction between Mip1p and Vps4p important for cytokinesis, and identify phosphorylation of Mid1p by aurora and polo kinases as being significant for this process.Author summaryReplication is a property of all living cells, with cell separation, so-called cytokinesis, the final step in the process. A large number of proteins have been identified that are required for cytokinesis, but in many cases it is not understand how they interact and regulate each other. In this research we have analysed two classes of proteins founds in all eukaryotic cells with central roles in cytokinesis: the endosomal sorting complex required for transport (ESCRT) proteins and the anillin protein Mid1p. We identify a direct physical interaction between the ESCRT protein Vps4 and anillin/Mid1p, and explore how it regulates cytokinesis. Midp1 activity is shown to controlled by the protein kinase Ark1p by direct phosphorylation, and this phosphorylation is important for Mid1p function. These observations identify new ways in which ESCRT and anillin/Mid1p control cell separation.

Development of a Human Symbiotic Assist Arm “PAS-Arm” (Experimental System and Creation of Virtual Guiding Surfaces)

We describe a new robotic assist device: the passive assist arm (PAS-Arm). PAS-Arms are intended for direct physical interaction with a human operator. PASArms are physically passive. The force to manipulate the arm end must be provided by the operator. Their purpose is not to enhance human strength, but to provide virtual guiding surfaces that constrain and guide the motion of the operator. PAS-Arms have three joints and a three dimensional workspace, but possess only two degrees of freedom due to the reduction of degrees of freedom created by a combination of Continuously Variable Transmissions (CVTs) and differential gears. In this paper, we first discuss the manipulability ellipsoid for the PAS-Arm. The major axis of the ellipsoid is the direction in which the arm end may be easily manipulated, and vice versa. We have developed an experimental system for the PAS-Arm. The CVTs of the experimental system may not adjust the transmission ratio to zero. Second, we describe an algorithm to address that problem. Finally, we present initial experiments that verify the PAS-Arm mechanism. The experimental results successfully produced virtual guiding surfaces.1 1. This paper is the full translation from the transactions of JSME, Series C, Vol.76, No.763, pp. 611-618, 2010.