Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition
| Autor: | Hiroo Ogi, Katsunori Sugimoto, Greicy H. Goto, Everett K. Henry, Avik Ghosh, Sevil Zencir |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
ATM protein
substitution mutation Cell Cycle Proteins Ataxia Telangiectasia Mutated Proteins Gene mutation medicine.disease_cause MEC1 protein S cerevisiae Phosphatidylinositol 3-Kinases FATC domain Protein structure RAD53 protein S cerevisiae genetics protein tertiary structure phosphatidylinositol 3 kinase gene mutation telomere homeostasis Phosphorylation DNA Fungal telomere Mutation Southern blotting Nuclear Functions protein kinase Tel1 Cell Cycle Intracellular Signaling Peptides and Proteins protein domain protein kinase Articles Telomere Protein-Serine-Threonine Kinases unclassified drug enzyme activity DNA-Binding Proteins priority journal autophosphorylation TEL1 protein S cerevisiae Saccharomyces cerevisiae Proteins DNA damage Molecular Sequence Data Protein domain protein localization Saccharomyces cerevisiae Protein Serine-Threonine Kinases Biology Article DNA damage checkpoint cell cycle protein protein serine threonine kinase plasmid Saccharomyces cerevisiae protein medicine double stranded DNA break signal peptide controlled study Amino Acid Sequence Protein kinase A protein expression Molecular Biology checkpoint kinase 2 fungal DNA Point mutation DNA Cell Biology G2-M DNA damage checkpoint Molecular biology DNA binding protein protein phosphorylation truncation mutation Protein Structure Tertiary Checkpoint Kinase 2 molecular genetics metabolism DNA Damage |
| Zdroj: | Molecular Biology of the Cell |
| ISSN: | 1939-4586 1059-1524 |
| Popis: | The FATC domain of Tel1 is studied via introduction of substitution and truncation mutations. It is found to be required for localization to sites of DNA damage and is essential for phosphorylation of exogenous substrates but dispensable for the intrinsic kinase activity. Two large phosphatidylinositol 3-kinase–related protein kinases (PIKKs), ATM and ATR, play a central role in the DNA damage response pathway. PIKKs contain a highly conserved extreme C-terminus called the FRAP-ATM-TRRAP-C-terminal (FATC) domain. In budding yeast, ATM and ATR correspond to Tel1 and Mec1, respectively. In this study, we characterized functions of the FATC domain of Tel1 by introducing substitution or truncation mutations. One substitution mutation, termed tel1-21, and a truncation mutation, called tel1-ΔC, did not significantly affect the expression level. The tel1-21 mutation impaired the cellular response to DNA damage and conferred moderate telomere maintenance defect. In contrast, the tel1-ΔC mutation behaved like a null mutation, conferring defects in both DNA damage response and telomere maintenance. Tel1-21 protein localized to DNA ends as effectively as wild-type Tel1 protein, whereas Tel1-ΔC protein failed. Introduction of a hyperactive TEL1-hy mutation suppressed the tel1-21 mutation but not the tel1-ΔC mutation. In vitro analyses revealed that both Tel1-21 and Tel1-ΔC proteins undergo efficient autophosphorylation but exhibit decreased kinase activities toward the exogenous substrate protein, Rad53. Our results show that the FATC domain of Tel1 mediates localization to DNA ends and contributes to phosphorylation of target proteins. |
| Databáze: | OpenAIRE |
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