Discovery of genes involved in mitosis, cell division, cell wall integrity and chromosome segregation through construction of Schizosaccharomyces pombe deletion strains

Abstract

The fission yeast model system Schizosaccharomyces pombe is used to study fundamental biological processes. To continue to fill gaps in the S. pombe gene deletion collection, we constructed a set of 90 haploid gene deletion strains covering many previously uncharacterized genes. To begin to understand the function of these genes, we exposed this collection of strains to a battery of stress conditions. Using this information in combination with microscopy, proteomics and mini-chromosome loss assays, we identified genes involved in cell wall integrity, cytokinesis, chromosome segregation and DNA metabolism. This subset of nonessential gene deletions will add to the toolkits available for the study of biological processes in S. pombe.

Nanoscale architecture of the Schizosaccharomyces pombe contractile ring

“…This may result in weakening the linkage between the strata of the contractile ring ultimately leading to loss of ring integrity and cell viability, irrespective of any mechanistic contributions of these components to ring function. In support of this hypothesis, synthetic lethality has been observed in several cases when null alleles of these genes were combined ( Ren et al, 2015 ; Pinar et al, 2008 ; Roberts-Galbraith et al, 2009 ; Liu et al, 2016 ; Chen et al, 2016 ). Indeed in the complete absence of Cdc15 and its SH3 interaction network, F-actin rings can still form but quickly disintegrate ( Laporte et al, 2011 ), underscoring the stabilizing function of the intermediate zone.…”

Roles of the novel coiled-coil protein Rng10 in septum formation during fission yeast cytokinesis

“…It is also unknown how Rga7 and Rng10 regulate Bgs1 and Bgs4 levels and dynamics. Because Rho1 overexpression can rescue both rga7 Δ and rng10 Δ cells and both deletions have synthetic interactions with mutations in rho1 and rho1 regulators ( Table 2 ; Chen et al. , 2016 ), Rga7 and Rng10 may regulate Bgs1 and Bgs4 through Rho1 GTPase and its GEFs and GAPs.…”

Cdk1 phosphorylation of fission yeast paxillin inhibits its cytokinetic ring localization

“…GFP-Pxl1 was purified using a nanobody-based IP (Rothbauer et al, 2008) that was performed as previously described (Chen et al, 2016) except that EDTA was removed from all buffers. A 2 L culture of GFP-pxl1 nda3-KM311 (KGY18174) was grown in 4X YES media at 32°C to OD 0.3 and then shifted to 18°C for 6.5 hours.…”

“…Studies of model organisms such as Schizosaccharomyces pombe have provided substantial insight into this process (Bathe and Chang, 2010;Pollard, 2010;Willet et al, 2015b). Forward genetic screens identified the proteins involved in cytokinesis, which include both CR structural components and regulators (Nurse et al, 1976;Minet et al, 1979;Chang et al, 1996;Balasubramanian et al, 1998;Kim et al, 2010;Hayles et al, 2013;Chen et al, 2015;Chen et al, 2016), many of which are conserved in the budding yeast Saccharomyces cerevisiae and metazoan cells (reviewed in (Pollard and Wu, 2010)). Subsequent fluorescence imaging studies determined the timing of cytokinesis events relative to mitosis in S. pombe (Wu et al, 2003;Wu and Pollard, 2005;Wu et al, 2006).…”