The novel (1,3)β-D-glucan synthase catalytic subunit Bgs4p from fission yeast is essential during both cytokinesis and polarized growth

Abstract

[EN]Schizosaccharomyces pombe contains four putative (1,3)β-D-glucan synthase (GS) catalytic subunits, Bgs1p to Bgs4p. In this work, we cloned bgs4+ and show that Bgs4p is the only subunit 1) essential for maintaining cell integrity during both cytokinesis and polarized growth, and 2) found to be part of the GS enzyme. Here we show that bgs4+, cwg1+ (cwg1-1 shows reduced cell-wall β-glucan and GS catalytic activity) and orb11+ (orb11-59 is defective in cell morphogenesis) are the same gene. bgs4+ is essential during spore germination. bgs4+ shut-off produces cell lysis at growing poles and mainly at the septum prior to cytokinesis, suggesting that Bgs4p is essential for cell wall growth and for compensating an excess of cell wall degradation during cytokinesis. Shut-off and overexpression analysis suggest that 1) Bgs4p forms part of a GS catalytic multiprotein complex, and 2) Bgs4p-promoted cell-wall β-glucan alterations induce compensatory mechanisms from other Bgs subunits and (1,3)α-D-glucan synthase. Physiological localization studies showed that Bgs4p localizes to the growing ends, the medial ring and septum, and in each process of wall synthesis or remodeling that occurs during sexual differentiation: mating, zygote and spore formation, and spore germination. Bgs4p timing and requirements for proper positioning during cytokinesis and its localization pattern during spore maturation differ from those of Bgs1p. Bgs4p localizes overlapping the contractile ring once Bgs1p is present and a Calcofluor white-stained septum material is detected, suggesting that Bgs4p is involved in a late process of secondary or general septum synthesis. Unlike Bgs1p, Bgs4p needs the medial ring but not the Septation Initiation Network proteins to localize with the other septation components. Furthermore, Bgs4p localization depends on the polarity establishment proteins. Finally, F-actin is necessary for Bgs4p delocalization from and relocalization to the growing regions, but it is not needed for its stable maintenance at the growing sites, poles and septum. All these data show for the first time an essential role for a Bgs subunit in the synthesis of a (1,3)β-D-glucan necessary to preserve cell integrity when cell wall synthesis or repair are needed.