New mutations in the core Schizosaccharomyces pombe spindle pole body scaffold Ppc89 reveal separable functions in regulating cell division.
Centrosomes and spindle pole bodies (SPB) are important for mitotic spindle formation and also serve as signaling platforms. In the fission yeast Schizosaccharomyces pombe, genetic ablation and high-resolution imaging indicate that the ɑ-helical Ppc89 is central to SPB structure and function. Here, we developed and characterized conditional and truncation mutants of ppc89. Alleles with mutations in two predicted ɑ-helices near the C-terminus were specifically defective in anchoring Sid4, the scaffold for the septation initiation network (SIN), and proteins dependent on Sid4 (Cdc11, Dma1, Mto1 and Mto2). Artificial tethering of Sid4 to the SPB fully rescued these ppc89 mutants. Another ppc89 allele had mutations located throughout the coding region. While this mutant was also defective in Sid4 anchoring, it displayed additional defects including fragmented SPBs and forming and constricting a second cytokinetic ring in one daughter cell. These defects were shared with a ppc89 allele truncated of the most C-terminal predicted ɑ-helices that is still able to recruit Sid4 and the SIN. We conclude that Ppc89 not only tethers the SIN to the SPB but is also necessary for the integrity of the SPB and faithful coordination of cytokinesis with mitosis.
Hanna SM, Tavafoghi B, Chen JS, Howard I, Ren L, Willet AH, Gould KL