Yeast Dun1 Kinase Regulates Ribonucleotide Reductase Small Subunit Localization in Response to Iron Deficiency
Ribonucleotide reductase (RNR) is an essential iron-dependent enzyme that catalyzes deoxyribonucleotide synthesis in eukaryotes. Living organisms have developed multiple strategies to tightly modulate RNR function to avoid inadequate or unbalanced deoxyribonucleotide pools that cause DNA damage and genome instability. Yeast cells activate RNR in response to genotoxic stress and iron deficiency by facilitating redistribution of its small heterodimeric subunit Rnr2-Rnr4 from the nucleus to the cytoplasm, where it forms an active holoenzyme with large Rnr1 subunit. Dif1 protein inhibits RNR by promoting nuclear import of Rnr2-Rnr4. Upon DNA damage, Dif1 phosphorylation by the Dun1 checkpoint kinase and its subsequent degradation enhances RNR function. In this report, we demonstrate that Dun1 kinase triggers Rnr2-Rnr4 redistribution to the cytoplasm in response to iron deficiency. We show that Rnr2-Rnr4 relocalization by low iron requires Dun1 kinase activity and phosphorylation site Thr-380 in the Dun1 activation loop, but not the Dun1 forkhead-associated domain. By using different Dif1 mutant proteins, we uncover that Dun1 phosphorylates Dif1 Ser-104 and Thr-105 residues upon iron scarcity. We observe that the Dif1 phosphorylation pattern differs depending on the stimuli, which suggests different Dun1 activating pathways. Importantly, the Dif1-S104A/T105A mutant exhibits defects in nucleus-to-cytoplasm redistribution of Rnr2-Rnr4 by iron limitation. Taken together, these results reveal that, in response to iron starvation, Dun1 kinase phosphorylates Dif1 to stimulate Rnr2-Rnr4 relocalization to the cytoplasm and promote RNR function.
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Language: | English |
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American Society for Biochemistry and Molecular Biology
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Subjects: | Iron deficiency, Yeast, Saccharomyces cerevisiae, Ribonucleotide reductase, |
Online Access: | http://hdl.handle.net/10261/131910 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100003359 http://dx.doi.org/10.13039/100000002 |
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dig-iata-es-10261-1319102021-12-27T15:59:50Z Yeast Dun1 Kinase Regulates Ribonucleotide Reductase Small Subunit Localization in Response to Iron Deficiency Sanvisens, Nerea Romero, Antonia Zhang, Caiguo Wu, Xiaorong An, Xiuxiang Huang, Mingxia Puig, Sergi Ministerio de Economía y Competitividad (España) Generalitat Valenciana National Institutes of Health (US) Iron deficiency Yeast Saccharomyces cerevisiae Ribonucleotide reductase Ribonucleotide reductase (RNR) is an essential iron-dependent enzyme that catalyzes deoxyribonucleotide synthesis in eukaryotes. Living organisms have developed multiple strategies to tightly modulate RNR function to avoid inadequate or unbalanced deoxyribonucleotide pools that cause DNA damage and genome instability. Yeast cells activate RNR in response to genotoxic stress and iron deficiency by facilitating redistribution of its small heterodimeric subunit Rnr2-Rnr4 from the nucleus to the cytoplasm, where it forms an active holoenzyme with large Rnr1 subunit. Dif1 protein inhibits RNR by promoting nuclear import of Rnr2-Rnr4. Upon DNA damage, Dif1 phosphorylation by the Dun1 checkpoint kinase and its subsequent degradation enhances RNR function. In this report, we demonstrate that Dun1 kinase triggers Rnr2-Rnr4 redistribution to the cytoplasm in response to iron deficiency. We show that Rnr2-Rnr4 relocalization by low iron requires Dun1 kinase activity and phosphorylation site Thr-380 in the Dun1 activation loop, but not the Dun1 forkhead-associated domain. By using different Dif1 mutant proteins, we uncover that Dun1 phosphorylates Dif1 Ser-104 and Thr-105 residues upon iron scarcity. We observe that the Dif1 phosphorylation pattern differs depending on the stimuli, which suggests different Dun1 activating pathways. Importantly, the Dif1-S104A/T105A mutant exhibits defects in nucleus-to-cytoplasm redistribution of Rnr2-Rnr4 by iron limitation. Taken together, these results reveal that, in response to iron starvation, Dun1 kinase phosphorylates Dif1 to stimulate Rnr2-Rnr4 relocalization to the cytoplasm and promote RNR function. This work has been supported by a predoctoral fellowship from “Conselleria d'Educació de la Generalitat Valenciana” (to N. S.), a predoctoral fellowship from the Spanish Ministry of Economy and Competitiveness (to A. M. R.), Spanish Ministry of Economy and Competitiveness Grants AGL2011-29099 and BIO2014-56298-P (to S. P.), and National Institutes of Health Grant CA125574 (to M. H.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Peer reviewed 2016-03 artículo http://purl.org/coar/resource_type/c_6501 Journal of Biological Chemistry 291:9807-9817 (2016) http://hdl.handle.net/10261/131910 10.1074/jbc.M116.720862 1083-351X http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100003359 http://dx.doi.org/10.13039/100000002 26970775 en #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2014-56298-P Postprint http://dx.doi.org/10.1074/jbc.M116.720862 Sí open American Society for Biochemistry and Molecular Biology |
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Iron deficiency Yeast Saccharomyces cerevisiae Ribonucleotide reductase Iron deficiency Yeast Saccharomyces cerevisiae Ribonucleotide reductase |
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Iron deficiency Yeast Saccharomyces cerevisiae Ribonucleotide reductase Iron deficiency Yeast Saccharomyces cerevisiae Ribonucleotide reductase Sanvisens, Nerea Romero, Antonia Zhang, Caiguo Wu, Xiaorong An, Xiuxiang Huang, Mingxia Puig, Sergi Yeast Dun1 Kinase Regulates Ribonucleotide Reductase Small Subunit Localization in Response to Iron Deficiency |
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Ribonucleotide reductase (RNR) is an essential iron-dependent enzyme that catalyzes deoxyribonucleotide synthesis in eukaryotes. Living organisms have developed multiple strategies to tightly modulate RNR function to avoid inadequate or unbalanced deoxyribonucleotide pools that cause DNA damage and genome instability. Yeast cells activate RNR in response to genotoxic stress and iron deficiency by facilitating redistribution of its small heterodimeric subunit Rnr2-Rnr4 from the nucleus to the cytoplasm, where it forms an active holoenzyme with large Rnr1 subunit. Dif1 protein inhibits RNR by promoting nuclear import of Rnr2-Rnr4. Upon DNA damage, Dif1 phosphorylation by the Dun1 checkpoint kinase and its subsequent degradation enhances RNR function. In this report, we demonstrate that Dun1 kinase triggers Rnr2-Rnr4 redistribution to the cytoplasm in response to iron deficiency. We show that Rnr2-Rnr4 relocalization by low iron requires Dun1 kinase activity and phosphorylation site Thr-380 in the Dun1 activation loop, but not the Dun1 forkhead-associated domain. By using different Dif1 mutant proteins, we uncover that Dun1 phosphorylates Dif1 Ser-104 and Thr-105 residues upon iron scarcity. We observe that the Dif1 phosphorylation pattern differs depending on the stimuli, which suggests different Dun1 activating pathways. Importantly, the Dif1-S104A/T105A mutant exhibits defects in nucleus-to-cytoplasm redistribution of Rnr2-Rnr4 by iron limitation. Taken together, these results reveal that, in response to iron starvation, Dun1 kinase phosphorylates Dif1 to stimulate Rnr2-Rnr4 relocalization to the cytoplasm and promote RNR function. |
author2 |
Ministerio de Economía y Competitividad (España) |
author_facet |
Ministerio de Economía y Competitividad (España) Sanvisens, Nerea Romero, Antonia Zhang, Caiguo Wu, Xiaorong An, Xiuxiang Huang, Mingxia Puig, Sergi |
format |
artículo |
topic_facet |
Iron deficiency Yeast Saccharomyces cerevisiae Ribonucleotide reductase |
author |
Sanvisens, Nerea Romero, Antonia Zhang, Caiguo Wu, Xiaorong An, Xiuxiang Huang, Mingxia Puig, Sergi |
author_sort |
Sanvisens, Nerea |
title |
Yeast Dun1 Kinase Regulates Ribonucleotide Reductase Small Subunit Localization in Response to Iron Deficiency |
title_short |
Yeast Dun1 Kinase Regulates Ribonucleotide Reductase Small Subunit Localization in Response to Iron Deficiency |
title_full |
Yeast Dun1 Kinase Regulates Ribonucleotide Reductase Small Subunit Localization in Response to Iron Deficiency |
title_fullStr |
Yeast Dun1 Kinase Regulates Ribonucleotide Reductase Small Subunit Localization in Response to Iron Deficiency |
title_full_unstemmed |
Yeast Dun1 Kinase Regulates Ribonucleotide Reductase Small Subunit Localization in Response to Iron Deficiency |
title_sort |
yeast dun1 kinase regulates ribonucleotide reductase small subunit localization in response to iron deficiency |
publisher |
American Society for Biochemistry and Molecular Biology |
url |
http://hdl.handle.net/10261/131910 http://dx.doi.org/10.13039/501100003329 http://dx.doi.org/10.13039/501100003359 http://dx.doi.org/10.13039/100000002 |
work_keys_str_mv |
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1777670023858356224 |