Coordination between fission yeast glucan formation and growth requires a sphingolipase activity.
css1 mutants display a novel defect in Schizosaccharomyces pombe cell wall formation. The mutant cells are temperature-sensitive and accumulate large deposits of material that stain with calcofluor and aniline blue in their periplasmic space. Biochemical analyses of this material indicate that it consists of alpha- and beta-glucans in the same ratio as found in cell walls of wild-type S. pombe. Strikingly, the glucan deposits in css1 mutant cells do not affect their overall morphology. The cells remain rod shaped, and the thickness of their walls is unaltered. Css1p is an essential protein related to mammalian neutral sphingomyelinase and is responsible for the inositolphosphosphingolipid-phospholipase C activity observed in S. pombe membranes. Furthermore, expression of css1(+) can compensate for loss of ISC1, the enzyme responsible for this activity in Saccharomyces cerevisiae membranes. Css1p localizes to the entire plasma membrane and secretory pathway; a C-terminal fragment of Css1p, predicted to encode a single membrane-spanning segment, is sufficient to direct membrane localization of the heterologous protein, GFP. Our results predict the existence of an enzyme(s) or process(es) essential for the coordination of S. pombe cell wall formation and division that is, in turn, regulated by a sphingolipid metabolite.
Feoktistova
A
A
Howard Hughes Medical Institute, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Magnelli
P
P
Abeijon
C
C
Perez
P
P
Lester
R L
RL
Dickson
R C
RC
Gould
K L
KL
D000595:Amino Acid Sequence; D002455:Cell Division; D002462:Cell Membrane / Q000201:enzymology; D002473:Cell Wall / Q000378:metabolism; D003001:Cloning, Molecular; D000939:Epitopes; D017353:Gene Deletion; D005936:Glucans / Q000378:metabolism*; D015151:Immunoblotting; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D012568:Schizosaccharomyces / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013107:Sphingolipids / Q000378:metabolism; D013108:Sphingomyelin Phosphodiesterase / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology*; D013347:Subcellular Fractions; D013696:Temperature; D013997:Time Factors; D010738:Type C Phospholipases / Q000378:metabolism
D000939:Epitopes; D005936:Glucans; D011134:Polysaccharides; D013107:Sphingolipids; D010738:Type C Phospholipases; D013108:Sphingomyelin Phosphodiesterase