In Vitro Analysis of Tem1 GTPase Activity and Regulation by the Bfa1/Bub2 GAP.

Marco Geymonat, Adonis Spanos, Katrin Rittinger

Methods in molecular biology (Clifton, N.J.) 2017

filter terms:

Tem1 is a small GTPase that controls the mitotic progression of Saccharomyces cerevisiae through the Mitotic Exit Network. Tem1 activity is tightly controlled in mitosis by Bub2 and Bfa1 and is also regulated by the spindle orientation checkpoint that monitors the correct alignment of the mitotic spindle with the mother-daughter axis. In this chapter we describe the purification of Tem1, Bfa1, and Bub2 and a detailed radioactive filter-binding assay to study the nucleotide binding properties of Tem1 and the role of its regulators Bfa1 and Bub2.

Citation

Marco Geymonat, Adonis Spanos, Katrin Rittinger. In Vitro Analysis of Tem1 GTPase Activity and Regulation by the Bfa1/Bub2 GAP. Methods in molecular biology (Clifton, N.J.). 2017;1505:71-80