Plo1 phosphorylates Dam1 to promote chromosome bi-orientation in fission yeast

Abstract

SummaryThe fungal-specific heterodecameric outer kinetochore DASH complex facilitates the interaction of kinetochores with spindle microtubules. In budding yeast, where kinetochores bind a single microtubule, the DASH complex is essential, and phosphorylation of Dam1 by the Aurora kinase homologue, Ipl1, causes detachment of kinetochores from spindle microtubules. We demonstrate that in the distantly related fission yeast, where the DASH complex is not essential for viability and kinetochores bind multiple microtubules, Dam1 is instead phosphorylated on serine 143 by the Polo kinase homologue, Plo1, during prometaphase and metaphase. This phosphorylation site is conserved in most fungal Dam1 proteins, including budding yeast Dam1. We show that Dam1 phosphorylation by Plo1 is dispensable for DASH assembly and chromosome retrieval but instead aids tension-dependent chromosome bi-orientation.

Dual role for fungal-specific outer kinetochore proteins during cell cycle and development in Magnaporthe oryzae

“…Similarly, mutations in budding yeast motor proteins, such as Kar3 (kinesin-14 family protein, Klp2 in S. pombe), Cin8, Kip1 (kinesin-5 family protein) and Kip3 (kinesin-8 family protein, Klp5/Klp6 in S. pombe) involved in MT stability, also display changes in spindle length (Straight et al, 1998;Zeng et al, 1999). Further, Dam1 plays a role in correct kinetochore attachments and bi-orientation in budding as well as fission yeast, lack of which leads to activation of the spindle assembly checkpoint (SAC), delaying the onset of anaphase (Janke et al, 2002;Sanchez-Perez et al, 2005;Buttrick et al, 2012). Dam1 phosphorylation by frequency of invasive hyphae at 36-40 hpi on rice leaf sheaths in the dam1Δ compared to the WT strain from three independent experiments.…”

Making an effective switch at the kinetochore by phosphorylation and dephosphorylation

“…Phosphorylation of the Ser20 residue of Dam1 disrupts its binding to the Ndc80 complex (Tien, et al 2010) while phosphorylation of C-terminal residues compromises Dam1 oligomerization (Wang, et al 2007). Although phosphorylation-mediated regulation of Dam1 does not seem to be conserved in fission yeast (Buttrick, et al 2012), in higher eukaryotes Aurora B-mediated phosphorylation negatively regulates kinetochore-localization and microtubule binding of the Ska complex, which has very similar functions as the Dam1 complex despite no apparent structural similarity (Jeyaprakash, et al 2012). The Ska complex binds to the Ndc80 complex and supports kinetochore-microtubule attachment by providing affinity for curved protofilaments, which facilitates binding to depolymerizing microtubules (Chan, et al 2012; Schmidt, et al 2012).…”

Polo kinase recruitment via the constitutive centromere-associated network at the kinetochore elevates centromeric RNA

“…This observation contrasts with a forced Cdc5 localization to both Cse4, which caused a strong growth inhibition but did not accumulate cells at any specific stage of the cell cycle [ 27 ], and to the DAM1/DASH1 subunit Dad4, which accumulated cells later in the cell cycle, possibly in late anaphase and/or telophase ( S4 Fig ). Notably, in fission yeast, it has been reported that Polo kinase, Plo1, phosphorylates Dam1 during prometaphase and metaphase for faithful chromosome biorientation [ 59 ].…”

“…cycle, possibly in late anaphase and/or telophase (S4 Fig). Notably, in fission yeast, it has been reported that Polo kinase, Plo1, phosphorylates Dam1 during prometaphase and metaphase for faithful chromosome biorientation [59].…”