Cited by 300DNA Repair MechanismsFungal and yeast genetics researchCancer-related Molecular Pathways

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Spk1/Rad53 is regulated by Mec1-dependent protein phosphorylation in DNA replication and damage checkpoint pathways.

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Sun, Z, Fay, D S, Marini, F, Foiani, M, Stern, D F·1996·Genes & Development

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Abstract

SPK1/RAD53/MEC2/SAD1 of Saccharomyces cerevisiae encodes an essential protein kinase that is required for activation of replication-sensitive and DNA damage-sensitive checkpoint arrest. We have investigated the regulation of phosphorylation and kinase activity of Spk1p during the cell cycle and by conditions that activate checkpoint pathways. Phosphorylation of Spk1p is induced by treatment of cells with agents that damage DNA or interfere with DNA synthesis. Although only S- and G2-phase cdc mutants arrest with hyperphosphorylated Spk1p, damage-induced phosphorylation of Spk1p can occur in G1 and M as well. Hydroxyurea (HU) induces phosphorylation of kinase-defective forms of Spk1p, demonstrating that this regulated phosphorylation of Spk1p occurs in trans. HU-induced phosphorylation is associated with increased catalytic activity of Spk1p. Furthermore, overexpression of wild-type SPK1, but not checkpoint-defective alleles, delays progression through the G1/S boundary. Damage-dependent phosphorylation of Spk1p requires both MEC1 and MEC3, whereas MEC1 but not MEC3, is required for replication block-induced phosphorylation. These data support the model that Spk1p is an essential intermediate component in a signal transduction pathway coupling damage and checkpoint functions to cell cycle arrest. This regulation is mediated through a protein kinase cascade that potentially includes Mec1p and Tel1p as the upstream kinases.

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Authors: Sun, Z., Fay, D. S., Marini, F., Foiani, M., & Stern, D. F.
Affiliations: Yale University (US)
ORCID: Sun, Z
Fay, D S
Marini, F
Foiani, M
Stern, D F
Venue: Genes & Development
Publisher: Cold Spring Harbor Laboratory
Published: 1996-02-15
Volume: 10
Issue: 4
Pages: 395-406
ISSN: 0890-9369 (ISSN-L)
1549-5477
DOI: 10.1101/gad.10.4.395
OpenAlex: W2150688223
Cited by: 300
References: 52
Citations by year: 2025: 2 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 1 · 2020: 3 · 2019: 4 · 2018: 6 · 2017: 5 · 2016: 8 · 2015: 4 · 2014: 7 · 2013: 6 · 2012: 4
Type: posted content
Language: en
Open access: bronze
doi.org
Locations: Genes & Development (publishedVersion)
PubMed (publishedVersion)
Archivio Istituzionale della Ricerca (Universita Degli Studi Di Milano) (submittedVersion, other-oa)
Funders: American Cancer Society
National Heart, Lung, and Blood Institute (R01HL175156)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
Keywords: Mec1, Tel1, DNA damage checkpoint, DNA replication checkpoint, protein phosphorylation, checkpoint signaling, cell cycle arrest, G1/S transition, hydroxyurea, kinase cascade, Saccharomyces cerevisiae
MeSH terms: Alkaline Phosphatase, Cell Cycle, Cell Division, DNA Damage, DNA Replication, DNA, Fungal, Fungal Proteins, Genes, Fungal, Hydroxyurea, Methyl Methanesulfonate, Phosphorylation, Precipitin Tests, Protein Kinases, Saccharomyces cerevisiae, Temperature, Immunoblotting, Signal Transduction, Gene Expression Regulation, Fungal, Mutagenesis, Protein Serine-Threonine Kinases, Cell Cycle Proteins, Saccharomyces cerevisiae Proteins, Intracellular Signaling Peptides and Proteins, Checkpoint Kinase 2

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Spk1/Rad53 is regulated by Mec1-dependent protein phosphorylation in DNA replication and damage checkpoint pathways.

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