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Mpg1, a fission yeast protein required for proper septum structure, is involved in cell cycle progression through cell-size checkpoint

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Abstract

Using a yeast two-hybrid screen we isolated a gene from Schizosaccharomyces pombe which corresponds to the previously uncharacterized ORF SPCC1906.01. We have designated this gene as mpg1, based on the putative function of its product as a mannose-1-phosphatase guanyltransferase. Mpg1 shows strong similarity to other GDP-mannose-1-phosphate guanyltransferases involved in the maintenance of cell wall integrity and/or glycosylation. This homology, together with the protein’s localization pattern demonstrated in this work, strongly suggests that Mpg1 is involved in cell wall and septum synthesis. Moreover, cells lacking Mpg1 present a defect in glycosylation, are more sensitive to Lyticase, and show an aberrant septum structure from the start of its deposition, indicating that the Mpg1 function is necessary for the correct assembly of the septum. Interestingly, lack of Mpg1 clearly affects cell cycle progression: mpg1 null mutants arrest as septated and bi-nucleated 4C cells, without an actomyosin ring. Wee1 is required for the G2/M arrest induced in the absence of Mpg1, since the blockade is circumvented when Wee1 is inactivated. Wee1 is part of a cell-size checkpoint that prevents entry into mitosis before cells reach a critical size. The results presented in this work demonstrate that the G2/M arrest induced in the absence of Mpg1 is mediated by this cell size checkpoint, since oversized mutant cells enter mitosis. The mpg1 loss-of-function mutant, therefore, provides a good model in which to study how cells coordinate cell growth and cell division.

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Acknowledgements

We thank Juan Jiménez and his laboratory for their interest and collaboration in this project. We also thank Manuel Cifuentes for his help with the generation of anti-Mpg1 antibodies. We are grateful to Antonio Jiménez and Manuel Cifuentes for their help with electron microscopy, to Manuel Muñoz for discussions and helpful suggestions, and Lucía Cruzado for technical support. We thank Sergio Moreno for the S. pombe library, Avelino Bueno for the Δflp1 strain and Juan Jiménez for the wee1-50 and cdc25-22 strains. We thank Dr. Javier Ruiz-Albert and Carmen R. Beuzón for critical reading of the manuscript and many helpful suggestions. I. Donoso was supported by a Predoctoral Research Fellowship from the Ministerio Español de Educación. M. A. Sánchez-Durán was supported by a Predoctoral Research Fellowship from the Junta de Andalucía. This work was supported by the Comisión Interministerial de Ciencia y Tecnología, Grants AGL2001-1857-C04-01, Plan Andaluz de Investigación, Group CVI 264, Universidad de Málaga and Gentech

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  1. M. C. Muñoz-Centeno

    Present address: Departamento de Genética, Universidad de Sevilla, 41012, Sevilla, Spain

  2. R. R. Daga

    Present address: Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide, 41013, Sevilla, Spain

Authors and Affiliations

  1. Departamento de Biología Celular, Genética y Fisiología, Universidad de Málaga, Campus de Teatinos, 29071, Málaga, Spain

    I. Donoso, M. C. Muñoz-Centeno, M. A. Sànchez-Durán, A. Flores, R. R. Daga, C. M. Guevara & E. R. Bejarano

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  1. I. Donoso
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Communicated by E. Bejarano

I. Donoso and M.C. Muñoz-Centeno contributed equally to this work

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Donoso, I., Muñoz-Centeno, M.C., Sànchez-Durán, M.A. et al. Mpg1, a fission yeast protein required for proper septum structure, is involved in cell cycle progression through cell-size checkpoint. Mol Genet Genomics 274, 155–167 (2005). https://doi.org/10.1007/s00438-005-0005-8

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