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dc.contributor.authorWaraky, Ahmed*
dc.contributor.authorLin, Yingbo*
dc.contributor.authorWarsito, Dudi*
dc.contributor.authorHaglund, Felix*
dc.contributor.authorAleem, Eiman*
dc.contributor.authorLarsson, Olle*
dc.date.accessioned2017-12-04T23:54:22Z
dc.date.available2017-12-04T23:54:22Z
dc.date.issued2017-11-03
dc.identifier.citationNuclear insulin-like growth factor 1 receptor phosphorylates proliferating cell nuclear antigen and rescues stalled replication forks after DNA damage 2017, 292 (44):18227 Journal of Biological Chemistryen
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.pmid28924044
dc.identifier.doi10.1074/jbc.M117.781492
dc.identifier.urihttp://hdl.handle.net/10150/626186
dc.description.abstractWe have previously shown that the insulin-like growth factor 1 receptor (IGF-1R) translocates to the cell nucleus, where it binds to enhancer-like regions and increases gene transcription. Further studies have demonstrated that nuclear IGF-1R (nIGF-1R) physically and functionally interacts with some nuclear proteins, i.e. the lymphoid enhancer-binding factor 1 (Lef1), histone H3, and Brahma-related gene-1 proteins. In this study, we identified the proliferating cell nuclear antigen (PCNA) as a nIGF-1R-binding partner. PCNA is a pivotal component of the replication fork machinery and a main regulator of the DNA damage tolerance (DDT) pathway. We found that IGF-1R interacts with and phosphorylates PCNA in human embryonic stem cells and other cell lines. In vitro MS analysis of PCNA co-incubated with the IGF-1R kinase indicated tyrosine residues 60, 133, and 250 in PCNA as IGF-1R targets, and PCNA phosphorylation was followed by mono- and polyubiquitination. Co-immunoprecipitation experiments suggested that these ubiquitination events may be mediated by DDT-dependent E2/E3 ligases (e.g. RAD18 and SHPRH/HLTF). Absence of IGF-1R or mutation of Tyr-60, Tyr-133, or Tyr-250 in PCNA abrogated its ubiquitination. Unlike in cells expressing IGF-1R, externally induced DNA damage in IGF-1R-negative cells caused G(1) cell cycle arrest and S phase fork stalling. Taken together, our results suggest a role of IGF-1R in DDT.
dc.description.sponsorshipSwedish Cancer Foundation; Swedish Research Council; Cancer Society in Stockholm; Swedish Children Cancer Society; Stockholm County Council; Karolinska Instituteten
dc.language.isoenen
dc.publisherAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCen
dc.relation.urlhttp://www.jbc.org/lookup/doi/10.1074/jbc.M117.781492en
dc.rights© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.en
dc.subjectinsulin-like growth factor (IGF)en
dc.subjectphosphorylationen
dc.subjectpost-transcriptional regulationen
dc.subjectproliferating cell nuclear antigen (PCNA)en
dc.subjectubiquitinen
dc.titleNuclear insulin-like growth factor 1 receptor phosphorylates proliferating cell nuclear antigen and rescues stalled replication forks after DNA damageen
dc.typeArticleen
dc.contributor.departmentUniv Arizona, Phoenix Childrens Hosp, Inst Mol Meden
dc.contributor.departmentUniv Arizona, Coll Med, Dept Child Hlthen
dc.identifier.journalJournal of Biological Chemistryen
dc.description.note12 month embargo; Published online: 18 September 2018en
dc.description.collectioninformationThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at repository@u.library.arizona.edu.en
dc.eprint.versionFinal published versionen
html.description.abstractWe have previously shown that the insulin-like growth factor 1 receptor (IGF-1R) translocates to the cell nucleus, where it binds to enhancer-like regions and increases gene transcription. Further studies have demonstrated that nuclear IGF-1R (nIGF-1R) physically and functionally interacts with some nuclear proteins, i.e. the lymphoid enhancer-binding factor 1 (Lef1), histone H3, and Brahma-related gene-1 proteins. In this study, we identified the proliferating cell nuclear antigen (PCNA) as a nIGF-1R-binding partner. PCNA is a pivotal component of the replication fork machinery and a main regulator of the DNA damage tolerance (DDT) pathway. We found that IGF-1R interacts with and phosphorylates PCNA in human embryonic stem cells and other cell lines. In vitro MS analysis of PCNA co-incubated with the IGF-1R kinase indicated tyrosine residues 60, 133, and 250 in PCNA as IGF-1R targets, and PCNA phosphorylation was followed by mono- and polyubiquitination. Co-immunoprecipitation experiments suggested that these ubiquitination events may be mediated by DDT-dependent E2/E3 ligases (e.g. RAD18 and SHPRH/HLTF). Absence of IGF-1R or mutation of Tyr-60, Tyr-133, or Tyr-250 in PCNA abrogated its ubiquitination. Unlike in cells expressing IGF-1R, externally induced DNA damage in IGF-1R-negative cells caused G(1) cell cycle arrest and S phase fork stalling. Taken together, our results suggest a role of IGF-1R in DDT.


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