Abstract
<p class="1Body">The heterodecameric Dam1 complex is involved in establishing and maintaining the connection between the kinetochore and the mitotic spindle during mitosis. Biochemical studies of the reconstituted complex have shed light upon how it interacts with microtubules. However, little information about the biochemical properties of the isolated subunits has been available. This report examines the stability and structure of Dad2p, one of the Dam1 complex subunits isolated from <em>Candida albicans</em>. By employing differential scanning fluorimetry, protease protection and hydrodynamic analyses, we show that Dad2p is specifically responsive to the presence of divalent cations. This observation may be important for understanding the dynamic structure and regulation of the Dam1 complex in fungal cells.</p>
Highlights
The process of mitosis has been studied extensively
This report examines the stability and structure of Dad2p, one of the Dam1 complex subunits isolated from Candida albicans
By employing differential scanning fluorimetry, protease protection and hydrodynamic analyses, we show that Dad2p is responsive to the presence of divalent cations
Summary
The process of mitosis has been studied extensively. While many fundamental mysteries remain to be unraveled, traditional biochemical and cell biological studies, as well as more recent genomic and proteomic approaches have resulted in a rather lengthy list of protein players necessary for correct chromosome segregation (Gascoigne & Cheeseman, 2011). This is the case in S. cerevisiae (Cheeseman et al, 2001; Janke, Ortiz, Tanaka, Lechner, & Schiebel, 2002; Li, Li, & Elledge, 2005) and C. albicans (Burrack, Applen, & Berman, 2011; Thakur & Sanyal, 2011), but not so for S. pombe (Sanchez-Perez et al, 2005) This difference has been postulated to reflect the more stringent requirement for maintaining connection with the depolymerizing microtubule when a single point of attachment is present; under these circumstances, the ring may provide necessary stability (Burrack et al, 2011; Thakur & Sanyal, 2011). This insight may have implications for future work in developing in vitro Dam complex reconstitution, for considering how the activity of the complex may be regulated in vivo, and for guiding the development of novel anti-fungal compounds
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