Fission yeast protein kinase C gene homologues are required for protoplast regeneration: A functional link between cell wall formation and cell shape control

Two novel protein kinase C (n PKC) gene homologues, pck1⁺ and pck2⁺ were isolated from the fission yeast Schizosaccharomyces pombe. We examined the functional differences of pck1⁺ and pck2⁺ in cell wall formation and actin organization of S. pombe. Regenerating protoplasts of a wild-type strain, single gene disruptants of pck1⁺ (Δpck1) and pck2⁺ (Δpck2) were used as a simple model to examine the functional links between PKC, cell wall formation and actin organization. Protoplasts of the wild type strain and those of Δpck1 reverted to intact cells in osmotically stabilized liquid medium. A close spatial association between new cell wall formation and actin was observed in these two strains. In Δpck2, protoplasts did not revert to intact cells: (1) scarcely any new cell wall material was formed; (2) actin was not reorganized; and (3) nuclear division and an increase in the amount of cytoplasm were observed in the regenerating protoplasts. These findings demonstrate that the pck2⁺ gene has a function essential for protoplast regeneration but the pck1⁺ gene does not. Involvement of n PKCs in cell wall formation and actin organization was also clarified. The effect of staurosporine (a potent inhibitor of protein kinases) on regenerating protoplasts of the three strains confirmed the assumption that the pck2 protein is an in vivo target of staurosporine in the fission yeast.