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Identification of protein kinase disruptions as suppressors of the calcium sensitivity of S. cerevisiae Δptp2 Δmsg5 protein phosphatase double disruptant

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Abstract

The double disruptant of the S. cerevisiae protein phosphatase (PPase) genes, PTP2 (phosphotyrosine-specific PPase) and MSG5 (phosphotyrosine and phosphothreonine/serine-PPase) causes calcium-sensitive growth (Cas). Previous study using Fluorescent-activated cell sorting (FACS) analysis showed that this growth defect with calcium occurs at G1–S transition in the cell cycle. We discovered that six non-essential protein kinase (PKase) disruptions (Δbck1, Δmkk1, Δslt2/Δmpk1, Δmck1, Δssk2 and Δyak1) suppressed the Cas-phenotype of the Δptp2 Δmsg5 double disruptant. Bck1p, Mkk1p and Slt2p are components of the mitogen-activated protein kinase (MAPK) cascade of cell wall integrity pathway (Slt2 pathway), and Mck1p is its down regulator. Ssk2p is the MAPK kinase kinase of the high-osmolarity glycerol (HOG) pathway, while Yak1p is a negative regulator for the cAMP-dependent PKA pathway. FACS analysis revealed that only the disruption of Δssk2 and Δyak1 but not Δbck1, Δmkk1, Δslt2 and Δmck1 was able to suppress the delayed G1–S transition, suggesting that suppression of the growth defect is not always accompanied by suppression of the G1–S transition delay. The discovery of these PKases as suppressors revealed that in addition to the previously anticipated Slt2 pathway, HOG, Yak1p and Mck1p regulatory pathways may also be involved in the calcium sensitivity of the Δptp2 Δmsg5 double disruptant.

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Abbreviations

PPase:

Protein phosphatase

PKase:

Protein kinase

MAPK:

Mitogen-activated protein kinase

Cas :

Calcium-sensitive growth

FACS:

Fluorescent-activated cell sorting

HOG:

High-osmolarity glycerol

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Acknowledgments

This work was supported by a Grant-in-Aid for Scientific Research B, 2007–2009, to S.H. from the Ministry of Education, Science, Sports and Culture of Japan.

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Authors and Affiliations

  1. Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Hermansyah, Walter A. Laviña, Minetaka Sugiyama, Yoshinobu Kaneko & Satoshi Harashima

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  1. Hermansyah
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  2. Walter A. Laviña
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  3. Minetaka Sugiyama
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  4. Yoshinobu Kaneko
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  5. Satoshi Harashima
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Correspondence to Satoshi Harashima.

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Communicated by Axel Brakhage.

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Hermansyah, Laviña, W.A., Sugiyama, M. et al. Identification of protein kinase disruptions as suppressors of the calcium sensitivity of S. cerevisiae Δptp2 Δmsg5 protein phosphatase double disruptant. Arch Microbiol 192, 157–165 (2010). https://doi.org/10.1007/s00203-009-0531-6

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  • DOI: https://doi.org/10.1007/s00203-009-0531-6

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