SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast

Barbara Baro, Soraya Játiva, Inés Calabria, Judith Vinaixa, Joan-Josep Bech-Serra, Carolina de LaTorre, João Rodrigues, María Luisa Hernáez, Concha Gil, Silvia Barceló-Batllori, Martin R Larsen, Ethel Queralt

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Resumé

Background: Protein phosphatase 2A (PP2A) is a family of conserved serine/threonine phosphatases involved in several essential aspects of cell growth and proliferation. PP2ACdc55 phosphatase has been extensively related to cell cycle events in budding yeast; however, few PP2ACdc55 substrates have been identified. Here, we performed a quantitative mass spectrometry approach to reveal new substrates of PP2ACdc55 phosphatase and new PP2A-related processes in mitotic arrested cells.

Results: We identified 62 statistically significant PP2ACdc55 substrates involved mainly in actin-cytoskeleton organization. In addition, we validated new PP2ACdc55 substrates such as Slk19 and Lte1, involved in early and late anaphase pathways, and Zeo1, a component of the cell wall integrity pathway. Finally, we constructed docking models of Cdc55 and its substrate Mob1. We found that the predominant interface on Cdc55 is mediated by a protruding loop consisting of residues 84-90, thus highlighting the relevance of these aminoacids for substrate interaction.

Conclusions: We used phosphoproteomics of Cdc55-deficient cells to uncover new PP2ACdc55 substrates and functions in mitosis. As expected, several hyperphosphorylated proteins corresponded to Cdk1-dependent substrates, although other kinases' consensus motifs were also enriched in our dataset, suggesting that PP2ACdc55 counteracts and regulates other kinases distinct from Cdk1. Indeed, Pkc1 emerged as a novel node of PP2ACdc55 regulation, highlighting a major role of PP2ACdc55 in actin cytoskeleton and cytokinesis, gene ontology terms significantly enriched in the PP2ACdc55-dependent phosphoproteome.

OriginalsprogEngelsk
Artikelnummergiy047
TidsskriftGigaScience
Vol/bind7
Udgave nummer5
Antal sider18
ISSN2047-217X
DOI
StatusUdgivet - 2018

Fingeraftryk

Phosphatases
Phosphoric Monoester Hydrolases
Yeast
CDC2 Protein Kinase
Proteins
Gene Ontology
Cytokinesis
Phosphoprotein Phosphatases
Substrates
Cell Wall
Cell Cycle
Cell Proliferation
Growth
Cells
Cell proliferation
Cell growth
Mass spectrometry
Ontology
Genes
Datasets

Citer dette

Baro, B., Játiva, S., Calabria, I., Vinaixa, J., Bech-Serra, J-J., de LaTorre, C., ... Queralt, E. (2018). SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast. GigaScience, 7(5), [giy047]. https://doi.org/10.1093/gigascience/giy047
Baro, Barbara ; Játiva, Soraya ; Calabria, Inés ; Vinaixa, Judith ; Bech-Serra, Joan-Josep ; de LaTorre, Carolina ; Rodrigues, João ; Hernáez, María Luisa ; Gil, Concha ; Barceló-Batllori, Silvia ; Larsen, Martin R ; Queralt, Ethel. / SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast. I: GigaScience. 2018 ; Bind 7, Nr. 5.
@article{a4e4ea4ef2e1456da40fcbbecedd69a4,
title = "SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast",
abstract = "Background: Protein phosphatase 2A (PP2A) is a family of conserved serine/threonine phosphatases involved in several essential aspects of cell growth and proliferation. PP2ACdc55 phosphatase has been extensively related to cell cycle events in budding yeast; however, few PP2ACdc55 substrates have been identified. Here, we performed a quantitative mass spectrometry approach to reveal new substrates of PP2ACdc55 phosphatase and new PP2A-related processes in mitotic arrested cells.Results: We identified 62 statistically significant PP2ACdc55 substrates involved mainly in actin-cytoskeleton organization. In addition, we validated new PP2ACdc55 substrates such as Slk19 and Lte1, involved in early and late anaphase pathways, and Zeo1, a component of the cell wall integrity pathway. Finally, we constructed docking models of Cdc55 and its substrate Mob1. We found that the predominant interface on Cdc55 is mediated by a protruding loop consisting of residues 84-90, thus highlighting the relevance of these aminoacids for substrate interaction.Conclusions: We used phosphoproteomics of Cdc55-deficient cells to uncover new PP2ACdc55 substrates and functions in mitosis. As expected, several hyperphosphorylated proteins corresponded to Cdk1-dependent substrates, although other kinases' consensus motifs were also enriched in our dataset, suggesting that PP2ACdc55 counteracts and regulates other kinases distinct from Cdk1. Indeed, Pkc1 emerged as a novel node of PP2ACdc55 regulation, highlighting a major role of PP2ACdc55 in actin cytoskeleton and cytokinesis, gene ontology terms significantly enriched in the PP2ACdc55-dependent phosphoproteome.",
keywords = "Mitosis, Mitotic exit network (MEN), Mob1, PP2A phosphatase, Phosphoproteomics, Pkc1, SILAC",
author = "Barbara Baro and Soraya J{\'a}tiva and In{\'e}s Calabria and Judith Vinaixa and Joan-Josep Bech-Serra and {de LaTorre}, Carolina and Jo{\~a}o Rodrigues and Hern{\'a}ez, {Mar{\'i}a Luisa} and Concha Gil and Silvia Barcel{\'o}-Batllori and Larsen, {Martin R} and Ethel Queralt",
year = "2018",
doi = "10.1093/gigascience/giy047",
language = "English",
volume = "7",
journal = "GigaScience",
issn = "2047-217X",
publisher = "BioMed Central",
number = "5",

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Baro, B, Játiva, S, Calabria, I, Vinaixa, J, Bech-Serra, J-J, de LaTorre, C, Rodrigues, J, Hernáez, ML, Gil, C, Barceló-Batllori, S, Larsen, MR & Queralt, E 2018, 'SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast', GigaScience, bind 7, nr. 5, giy047. https://doi.org/10.1093/gigascience/giy047

SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast. / Baro, Barbara; Játiva, Soraya; Calabria, Inés; Vinaixa, Judith; Bech-Serra, Joan-Josep; de LaTorre, Carolina; Rodrigues, João; Hernáez, María Luisa; Gil, Concha; Barceló-Batllori, Silvia; Larsen, Martin R; Queralt, Ethel.

I: GigaScience, Bind 7, Nr. 5, giy047, 2018.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast

AU - Baro, Barbara

AU - Játiva, Soraya

AU - Calabria, Inés

AU - Vinaixa, Judith

AU - Bech-Serra, Joan-Josep

AU - de LaTorre, Carolina

AU - Rodrigues, João

AU - Hernáez, María Luisa

AU - Gil, Concha

AU - Barceló-Batllori, Silvia

AU - Larsen, Martin R

AU - Queralt, Ethel

PY - 2018

Y1 - 2018

N2 - Background: Protein phosphatase 2A (PP2A) is a family of conserved serine/threonine phosphatases involved in several essential aspects of cell growth and proliferation. PP2ACdc55 phosphatase has been extensively related to cell cycle events in budding yeast; however, few PP2ACdc55 substrates have been identified. Here, we performed a quantitative mass spectrometry approach to reveal new substrates of PP2ACdc55 phosphatase and new PP2A-related processes in mitotic arrested cells.Results: We identified 62 statistically significant PP2ACdc55 substrates involved mainly in actin-cytoskeleton organization. In addition, we validated new PP2ACdc55 substrates such as Slk19 and Lte1, involved in early and late anaphase pathways, and Zeo1, a component of the cell wall integrity pathway. Finally, we constructed docking models of Cdc55 and its substrate Mob1. We found that the predominant interface on Cdc55 is mediated by a protruding loop consisting of residues 84-90, thus highlighting the relevance of these aminoacids for substrate interaction.Conclusions: We used phosphoproteomics of Cdc55-deficient cells to uncover new PP2ACdc55 substrates and functions in mitosis. As expected, several hyperphosphorylated proteins corresponded to Cdk1-dependent substrates, although other kinases' consensus motifs were also enriched in our dataset, suggesting that PP2ACdc55 counteracts and regulates other kinases distinct from Cdk1. Indeed, Pkc1 emerged as a novel node of PP2ACdc55 regulation, highlighting a major role of PP2ACdc55 in actin cytoskeleton and cytokinesis, gene ontology terms significantly enriched in the PP2ACdc55-dependent phosphoproteome.

AB - Background: Protein phosphatase 2A (PP2A) is a family of conserved serine/threonine phosphatases involved in several essential aspects of cell growth and proliferation. PP2ACdc55 phosphatase has been extensively related to cell cycle events in budding yeast; however, few PP2ACdc55 substrates have been identified. Here, we performed a quantitative mass spectrometry approach to reveal new substrates of PP2ACdc55 phosphatase and new PP2A-related processes in mitotic arrested cells.Results: We identified 62 statistically significant PP2ACdc55 substrates involved mainly in actin-cytoskeleton organization. In addition, we validated new PP2ACdc55 substrates such as Slk19 and Lte1, involved in early and late anaphase pathways, and Zeo1, a component of the cell wall integrity pathway. Finally, we constructed docking models of Cdc55 and its substrate Mob1. We found that the predominant interface on Cdc55 is mediated by a protruding loop consisting of residues 84-90, thus highlighting the relevance of these aminoacids for substrate interaction.Conclusions: We used phosphoproteomics of Cdc55-deficient cells to uncover new PP2ACdc55 substrates and functions in mitosis. As expected, several hyperphosphorylated proteins corresponded to Cdk1-dependent substrates, although other kinases' consensus motifs were also enriched in our dataset, suggesting that PP2ACdc55 counteracts and regulates other kinases distinct from Cdk1. Indeed, Pkc1 emerged as a novel node of PP2ACdc55 regulation, highlighting a major role of PP2ACdc55 in actin cytoskeleton and cytokinesis, gene ontology terms significantly enriched in the PP2ACdc55-dependent phosphoproteome.

KW - Mitosis

KW - Mitotic exit network (MEN)

KW - Mob1

KW - PP2A phosphatase

KW - Phosphoproteomics

KW - Pkc1

KW - SILAC

U2 - 10.1093/gigascience/giy047

DO - 10.1093/gigascience/giy047

M3 - Journal article

C2 - 29688323

VL - 7

JO - GigaScience

JF - GigaScience

SN - 2047-217X

IS - 5

M1 - giy047

ER -

Baro B, Játiva S, Calabria I, Vinaixa J, Bech-Serra J-J, de LaTorre C et al. SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast. GigaScience. 2018;7(5). giy047. https://doi.org/10.1093/gigascience/giy047