Regulation of tolerance to DNA alkylating damage by Dot1 and Rad53 in Saccharomyces cerevisiae
| Autor: | David Ontoso, Alfonso Gallego-Sánchez, Isabel Acosta, Pedro A. San-Segundo, Avelino Bueno, Francisco Conde |
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| Rok vydání: | 2010 |
| Předmět: |
DNA Replication
Alkylating Agents Saccharomyces cerevisiae Proteins DNA Repair DNA repair DNA damage Cell Cycle Proteins Saccharomyces cerevisiae Protein Serine-Threonine Kinases Biochemistry Ionizing radiations Histones chemistry.chemical_compound TLS PCNA Phosphorylation DNA Fungal Molecular Biology biology DNA replication Nuclear Proteins Cell Biology Histone-Lysine N-Methyltransferase Methyl Methanesulfonate Molecular biology MMS Chromatin Methyl methanesulfonate Proliferating cell nuclear antigen Checkpoint Kinase 2 Histone chemistry Mutagenesis biology.protein IR Histone Methyltransferases REV1 Methylmethane sulfonate DNA Damage |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 1568-7856 |
| Popis: | 12 páginas, 8 figuras, 2 tablas.-- El pdf del artículo es la versión de autor. To maintain genomic integrity cells have to respond properly to a variety of exogenous and endogenous factors that produce genome injuries and interfere with DNA replication. DNA integrity checkpoints coordinate this response by slowing cell cycle progression to provide time for the cell to repair the damage, stabilizing replication forks and stimulating DNA repair to restore the original DNA sequence and structure. In addition, there are also mechanisms of damage tolerance, such as translesion synthesis (TLS), which are important for survival after DNA damage. TLS allows replication to continue without removing the damage, but results in a higher frequency of mutagenesis. Here, we investigate the functional contribution of the Dot1 histone methyltransferase and the Rad53 checkpoint kinase to TLS regulation in Saccharomyces cerevisiae. We demonstrate that the Dot1-dependent status of H3K79 methylation modulates the resistance to the alkylating agent MMS, which depends on PCNA ubiquitylation at lysine 164. Strikingkly, either the absence of DOT1, which prevents full activation of Rad53, or the expression of an HA-tagged version of RAD53, which produces low amounts of the kinase, confer increased MMS resistance. However, the dot1Δ rad53-HA double mutant is hypersensitive to MMS and shows barely detectable amounts of activated kinase. Furthermore, moderate overexpression of RAD53 partially suppresses the MMS resistance of dot1Δ. In addition, we show that MMS-treated dot1Δ and rad53-HA cells display increased number of chromosome-associated Rev1 foci. We propose that threshold levels of Rad53 activity exquisitely modulate the tolerance to alkylating damage at least by controlling the abundance of the key TLS factor Rev1 bound to chromatin. DO and AGS were supported by predoctoral fellowships from CSIC and MEC (Spain), respectively. Research in our labs is supported by grants from Ministry of Science and Innovation of Spain (BFU2009-06938 to AB and BFU2009-07159 to PSS) and a grant from Fundación Ramón Areces to PSS. |
| Databáze: | OpenAIRE |
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