Ras/cAMP-dependent Protein Kinase (PKA) Regulates Multiple Aspects of Cellular Events by Phosphorylating the Whi3 Cell Cycle Regulator in Budding Yeast

Mizunuma, Masaki; Tsubakiyama, Ryohei; Ogawa, Takafumi; Shitamukai, Atsunori; Kobayashi, Yoshifumi; Inai, Tomomi; Kume, Kazunori; Hirata, Dai

doi:10.1074/jbc.m112.402214

Cited by 25 publications

(32 citation statements)

References 21 publications

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“…Phosphorylation of Whi3 is regulated by the Ras/cAMP PKA pathway. The interaction between phosphorylated Whi3 and the CLN3 G1 cyclin messenger RNA could accelerate the G1/S transition (Mizunuma et al, ). In this study, Baf‐A1 inhibited the activity of V‐ATPase, which may promote the transition from G1 phase to the S phase by affecting the PKA pathway and phosphorylation of Whi3 in BmE cells.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of silkworm vacuolar‐type ATPase activity by its inhibitor Bafilomycin A1 induces caspase‐dependent apoptosis in an embryonic cell line of silkworm

Tan

Wang

Liu

et al. 2018

Arch Insect Biochem Physiol

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Vacuolar‐type ATPase (V‐ATPase) is a type of hydrogen ion transporter located in the vesicular membrane‐like system, which mediates active transport and intracellular acidification in various compartments. In mammals, V‐ATPase has been reported to play a key role in cell proliferation and apoptosis. The studies of V‐ATPase in silkworm mainly focus on the acidification regulation of midgut and silk gland and immune resistance. However, there are few reports about the function of silkworm V‐ATPase on cell proliferation, autophagy, and apoptosis. Thus, the function of V‐ATPase in a cell line of Bombyx mori (BmE) was investigated by treating the cell line with bafilomycin A1, a specific inhibitor of V‐ATPase. Cell counting kit 8 (CCK8) and flow cytometry analysis showed that bafilomycin A1 treatment decreased the cell proliferation activity, affected the cell cycle progression and induced cell apoptosis. LysoTracker Red staining showed that the target of bafilomycin A1 is lysosome. The expression of all autophagy‐related genes ( BmATG5, BmATG6, and BmATG8) decreased, indicating that cell autophagy was inhibited. The analysis of the apoptosis pathway demonstrated that inhibiting the activity of V‐ATPase of BmE cells could promote mitochondria to release cytochrome C, inhibit the expression of BmIAP, and activate the caspase cascade to induce apoptosis. All these findings systematically illustrate the effects of V‐ATPase on the proliferation, autophagy, and apoptosis in BmE cells, and provide new ideas and a theoretical basis for further study on the function of V‐ATPase in BmE.

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Section: Discussionmentioning

confidence: 99%

Inhibition of silkworm vacuolar‐type ATPase activity by its inhibitor Bafilomycin A1 induces caspase‐dependent apoptosis in an embryonic cell line of silkworm

Tan

Wang

Liu

et al. 2018

Arch Insect Biochem Physiol

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“…The molecular mechanisms that determine whether a cell will execute entry into the cell cycle from G1—also known as S tart in yeast—have been well characterized. In early G1, the only available cyclin is Cln3, whose rate of synthesis is proportional to cell mass ( Hall et al ., 1998 ; Nash et al ., 2001 ; Alberghina et al ., 2004 ; Mizunuma et al ., 2013 ; Yahya et al ., 2014 ). Moreover, Cln3 is sequestered at the endoplasmic reticulum (ER) by Whi3 in an Ssa1-dependent process ( Gari et al ., 2001 ; Ferrezuelo et al ., 2012 ; Caudron and Barral, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Experimental testing of a new integrated model of the budding yeast Starttransition

Adames

Schuck

Chen

et al. 2015

MBoC

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“…In Saccharomyces cerevisiae, Whi3 has been seen to localize to stress granules and P bodies following certain stress conditions (Cai et al, 2013;Mizunuma et al, 2013). We hypothesize that a subset of hyphal Whi3 puncta may be used to regulate other Whi3 phenotypes related to environmental stress response, and S283 might be involved in the regulation of assemblies not seen under normal conditions.…”

Section: Whi3 Residues Phosphorylated During Heat Stress Do Not Affecmentioning

confidence: 82%

“…The second separation of function allele we identified was S637A. This site is located within the RRM of the protein and is a conserved, putative PKA consensus site that has been shown experimentally to be phosphorylated in Saccharomyces cerevisiae (Mizunuma et al, 2013). The S637A mutant has normal nuclear asynchrony ( Fig.…”

Section: Separation Of Function Mutants Specific For Asynchrony or Cementioning

confidence: 87%

Phosphoregulation provides functional specificity to biomolecular condensates in the cell cycle and cell polarity

Gerbich

Gladfelter²

2019

Preprint

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words)Residue specific phosphorylation of the RNA-binding protein Whi3 is used to specifically regulate subsets of functionally distinct condensates in the multinucleate fungus Ashbya gossypii. AbstractCytoplasmic patterning is a feature of many cells from embryos to neurons and fungi. Biomolecular condensation is a way of organizing cytosol in which proteins and nucleic acids coassemble into compartments. How molecular identity of condensates is achieved is not well understood. In the multinucleate, filamentous fungus Ashbya gossypii, the RNA-binding protein Whi3 regulates the cell cycle and cell polarity through forming macromolecular structures that behave like condensates. Whi3 has distinct spatial localizations and mRNA targets making it a powerful model for how, when and where specific identities are established for condensates. Using mass-spectrometry, we identified residues on Whi3 that are differentially phosphorylated under specific conditions and generated mutants which ablate this regulation. This yielded separation of function alleles that were functional for either cell polarity or nuclear cycling but not both. This study shows that phosphorylation of individual residues on molecules in biomolecular condensates can provide specificity that give rise to distinct functional identities in the same cell.

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Ras/cAMP-dependent Protein Kinase (PKA) Regulates Multiple Aspects of Cellular Events by Phosphorylating the Whi3 Cell Cycle Regulator in Budding Yeast

Cited by 25 publications

References 21 publications

Inhibition of silkworm vacuolar‐type ATPase activity by its inhibitor Bafilomycin A1 induces caspase‐dependent apoptosis in an embryonic cell line of silkworm

Inhibition of silkworm vacuolar‐type ATPase activity by its inhibitor Bafilomycin A1 induces caspase‐dependent apoptosis in an embryonic cell line of silkworm

Experimental testing of a new integrated model of the budding yeast Starttransition

Phosphoregulation provides functional specificity to biomolecular condensates in the cell cycle and cell polarity

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