A Coordinated Interdependent Protein Circuitry Stabilizes the Kinetochore Ensemble to Protect CENP-A in the Human Pathogenic Yeast Candida albicans
Figure 2
Stability of the kinetochore structure is independent of structural integrity of the mitotic spindle.
A) Wild-type BWP17 cells, untreated (DMSO) or treated with nocodazole (+NOC), were fixed and stained with DAPI, and anti-tubulin antibodies. As compared to untreated cells (upper panels), NOC treated cells (lower panels) exhibited a significant loss of tubulin staining indicating a compromised structure of the mitotic spindle as expected by NOC treatment. (B) CENP-A/Cse4-GFP signals were analyzed in absence (DMSO) or presence of NOC (+NOC) in 10118 (CSE4:GFP:CSE4/cse4) cells. (C) Similarly, Mtw1-GFP signals were examined in absence (DMSO) or presence of NOC in YJB10695 (MTW1GFP/MTW1) cells. No significant change in intensity of either Cse4-GFP or Mtw1-GFP signals in NOC treated cells. (D) PCR analysis on Cse4 ChIP-DNA obtained from NOC treated BWP17 cells. Primers from CENR (CaChrR: 1747812–1748023), CEN1 (CaChr1: 1565723–1565967), CEN7 (CaChr7: 427369–427560) or 17 kb away from CEN7 (CaChr7: 444584–444875) (non-CEN) were used for PCR analysis. T, total DNA; +, IP DNA with anti-Cse4 antibodies and −, beads only control without antibodies. Bars, 5 µm.