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

1994 ◽  
Vol 107 (5) ◽  
pp. 1131-1136 ◽  
Author(s):  
H. Kobori ◽  
T. Toda ◽  
H. Yaguchi ◽  
M. Toya ◽  
M. Yanagida ◽  
...  
Keyword(s):  
Cell Wall ◽  
Fission Yeast ◽  
Wild Type Strain ◽  
Protoplast Regeneration ◽  
Cell Wall Formation ◽  
Wild Type ◽  
Intact Cells ◽  
Actin Organization ◽  
Wall Formation ◽  
Kinase C

Two novel protein kinase C (n PKC) gene homologues, pck1+ and pck2+ were isolated from the fission yeast Schizosaccharomyces pombe (Toda et al. (1993) EMBO J. 12, 1987). 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+ (delta pck1) and pck2+ (delta pck2) 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 delta pck1 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 delta pck2, 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.

scholarly journals Coordination Between Fission Yeast Glucan Formation and Growth Requires a Sphingolipase Activity

Genetics ◽  
2001 ◽  
Vol 158 (4) ◽  
pp. 1397-1411 ◽  
Author(s):  
Anna Feoktistova ◽  
Paula Magnelli ◽  
Claudia Abeijon ◽  
Pilar Perez ◽  
Robert L Lester ◽  
...  
Keyword(s):  
Cell Wall ◽  
Secretory Pathway ◽  
Temperature Sensitive ◽  
Cell Wall Formation ◽  
Wild Type ◽  
Wall Formation ◽  
Biochemical Analyses ◽  
Membrane Spanning ◽  
Mutant Cells ◽  
Large Deposits

Abstractcss1 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 α- and β-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.

scholarly journals Establishment of an Efficient Protoplast Regeneration and Transfection Protocol for Field Cress (Lepidium campestre)

2021 ◽  
Vol 3 ◽  
Author(s):  
Sjur Sandgrind ◽  
Xueyuan Li ◽  
Emelie Ivarson ◽  
Annelie Ahlman ◽  
Li-Hua Zhu
Keyword(s):  
Cell Wall ◽  
Plant Species ◽  
Protoplast Regeneration ◽  
Cell Wall Formation ◽  
Wall Formation ◽  
Transfection Protocol ◽  
Culture Duration ◽  
Lepidium Campestre ◽  
Basal Media ◽  
Field Cress

Field cress (Lepidium campestre) is a potential oilseed crop that has been under domestication in recent decades. CRISPR/Cas9 is a powerful tool for rapid trait improvement and gene characterization and for generating transgene-free mutants using protoplast transfection system. However, protoplast regeneration remains challenging for many plant species. Here we report an efficient protoplast regeneration and transfection protocol for field cress. Important factors such as type of basal media, type/combination of plant growth regulators, and culture duration on different media were optimized. Among the basal media tested, Nitsch was the best for protoplast growth in MI and MII media. For cell wall formation during the early stage of protoplast growth, relatively high auxin concentrations (0.5 mg L−1 NAA and 2,4-D), without addition of cytokinin was preferred for maintaining protoplast viability. After cell wall formation, 1.1 mg L−1 TDZ combined with either 0.05 mg L−1 NAA or 2,4-D was found to efficiently promote protoplast growth. On solid shoot induction medium, 1.1 mg L−1 TDZ without any auxin resulted in over 80% shoot generation frequency. A longer culture duration in MI medium would inhibit protoplast growth, while a longer culture duration in MII medium significantly delayed shoot formation. Using this optimized protoplast regeneration protocol, we have established an efficient PEG-mediated transfection protocol using a vector harboring the GFP gene, with transfection efficiencies of 50–80%. This efficient protoplast protocol would facilitate further genetic improvement of field cress via genome editing, and be beneficial to development of protoplast regeneration protocols for related plant species.

Dynamics of Cell Wall Formation in Fission Yeast,Schizosaccharomyces pombe

1998 ◽  
Vol 24 (1-2) ◽  
pp. 178-206 ◽  
Author(s):  
Masako Osumi ◽  
Mamiko Sato ◽  
Sanae A Ishijima ◽  
Mami Konomi ◽  
Tomoko Takagi ◽  
...  
Keyword(s):  
Cell Wall ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Cell Wall Formation ◽  
Wall Formation

scholarly journals Genetical and structural analyses of cell-wall formation in Chlamydomonas reinhardi

1971 ◽  
Vol 17 (1) ◽  
pp. 33-43 ◽  
Author(s):  
D. Roy Davies ◽  
A. Plaskitt
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Single Gene ◽  
Three Dimensional ◽  
Cell Wall Formation ◽  
Wild Type ◽  
Nuclear Level ◽  
Wall Formation ◽  
Cell Wall Biogenesis ◽  
Mutant Strains

SUMMARYFifteen mutant strains of Chlamydomonas reinhardi were isolated which showed defects in some aspect of the process of cell-wall formation. Genetic analyses indicated that most of the mutations were due to single gene changes; two were anomalous in that non-Mendelian segregations were obtained on crossing with other genotypes, and on selfing they frequently gave rise to wild-type phenotypes.Occasional somatic revertants were also obtained from these two strains. On the basis of these analyses it is suggested that there are two levels of control operating in the process of cell wall biogenesis - one concerned with subunit production at the nuclear level and another, possibly concerned with three-dimensional organization, at another level. Electron-microscope analyses of the different mutants showed the mutants to be divided into three main categories: those in which the wall was formed but was not attached to the plasma membrane, those in which the wall was attached to the membrane, and those in which very little wall was formed. In the last class in particular, vesicles containing wall precursors were clearly visible, and were shed through the plasma membrane into the medium.

Ethylene effects on cambial activity and cell wall formation in hypocotyls of Picea abies seedlings

1991 ◽  
Vol 82 (2) ◽  
pp. 219-224 ◽  
Author(s):  
Barbro S. M. Ingemarsson ◽  
Leif Eklund ◽  
Lennart Eliasson
Keyword(s):  
Cell Wall ◽  
Picea Abies ◽  
Cambial Activity ◽  
Cell Wall Formation ◽  
Wall Formation

Arabidopsis Callose Synthase Gene GSL8 is Required for Cell Wall Formation and Establishment and Maintenance of Quiescent Center

2014 ◽  
Vol 48 (4) ◽  
pp. 389-397
Author(s):  
Liu Lin ◽  
Quan Xianqing ◽  
Zhao Xiaomei ◽  
Huang Lihua ◽  
Feng Shangcai ◽  
...  
Keyword(s):  
Cell Wall ◽  
Quiescent Center ◽  
Cell Wall Formation ◽  
Callose Synthase ◽  
Wall Formation
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