The KYxxL motif in Rad17 protein is essential for the interaction with the 9–1–1 complex

Fukumoto, Yasunori; Ikeuchi, Masayoshi; Nakayama, Yuji; Yamaguchi, Naoto

doi:10.1016/J.BBRC.2016.07.014

Title: The KYxxL motif in Rad17 protein is essential for the interaction with the 9–1–1 complex

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Abstract

ATR-dependent DNA damage checkpoint is the major DNA damage checkpoint against UV irradiation and DNA replication stress. The Rad17–RFC and Rad9–Rad1–Hus1 (9–1–1) complexes interact with each other to contribute to ATR signaling, however, the precise regulatory mechanism of the interaction has not been established. Here, we identified a conserved sequence motif, KYxxL, in the AAA+ domain of Rad17 protein, and demonstrated that this motif is essential for the interaction with the 9–1–1 complex. We also show that UV-induced Rad17 phosphorylation is increased in the Rad17 KYxxL mutants. These data indicate that the interaction with the 9–1–1 complex is not required for Rad17 protein to be an efficient substrate for the UV-induced phosphorylation. Our data also raise the possibility that the 9–1–1 complex plays a negative regulatory role in the Rad17 phosphorylation. We also show that the nucleotide-binding activity of Rad17 is required for its nuclear localization. - Highlights: • We have identified a conserved KYxxL motif in Rad17 protein. • The KYxxL motif is crucial for the interaction with the 9–1–1 complex. • The KYxxL motif is dispensable or inhibitory for UV-induced Rad17 phosphorylation. • Nucleotide binding of Rad17 is required for its nuclear localization.

Authors:

Fukumoto, Yasunori ^[1]; Ikeuchi, Masayoshi; Nakayama, Yuji ^[2]; Yamaguchi, Naoto ^[1]

Laboratory of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675 (Japan)
Department of Biochemistry & Molecular Biology, Kyoto Pharmaceutical University, Kyoto 607-8414 (Japan)

Publication Date:: Fri Sep 02 00:00:00 EDT 2016

OSTI Identifier:: 22606196

Resource Type:: Journal Article

Journal Name:: Biochemical and Biophysical Research Communications

Additional Journal Information:: Journal Volume: 477; Journal Issue: 4; Other Information: Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X

Country of Publication:: United States

Language:: English

Subject:: 60 APPLIED LIFE SCIENCES; AMINO ACIDS; ANTIGENS; CELL CYCLE; COMPLEXES; DNA; DNA DAMAGES; DNA REPLICATION; IRRADIATION; MUTANTS; NUCLEOTIDES; PHOSPHORYLATION; PROTEINS; ULTRAVIOLET RADIATION

Citation Formats


                    Fukumoto, Yasunori, Ikeuchi, Masayoshi, Nakayama, Yuji, and Yamaguchi, Naoto. The KYxxL motif in Rad17 protein is essential for the interaction with the 9–1–1 complex.  United States: N. p., 2016. 
        Web.  doi:10.1016/J.BBRC.2016.07.014.

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                    Fukumoto, Yasunori, Ikeuchi, Masayoshi, Nakayama, Yuji, & Yamaguchi, Naoto. The KYxxL motif in Rad17 protein is essential for the interaction with the 9–1–1 complex.  United States.  https://doi.org/10.1016/J.BBRC.2016.07.014

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                    Fukumoto, Yasunori, Ikeuchi, Masayoshi, Nakayama, Yuji, and Yamaguchi, Naoto. 2016.  
        "The KYxxL motif in Rad17 protein is essential for the interaction with the 9–1–1 complex".  United States.  https://doi.org/10.1016/J.BBRC.2016.07.014.

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@article{osti_22606196,

  title        = {The KYxxL motif in Rad17 protein is essential for the interaction with the 9–1–1 complex},

  author       = {Fukumoto, Yasunori and Ikeuchi, Masayoshi and Nakayama, Yuji and Yamaguchi, Naoto},

  abstractNote = {ATR-dependent DNA damage checkpoint is the major DNA damage checkpoint against UV irradiation and DNA replication stress. The Rad17–RFC and Rad9–Rad1–Hus1 (9–1–1) complexes interact with each other to contribute to ATR signaling, however, the precise regulatory mechanism of the interaction has not been established. Here, we identified a conserved sequence motif, KYxxL, in the AAA+ domain of Rad17 protein, and demonstrated that this motif is essential for the interaction with the 9–1–1 complex. We also show that UV-induced Rad17 phosphorylation is increased in the Rad17 KYxxL mutants. These data indicate that the interaction with the 9–1–1 complex is not required for Rad17 protein to be an efficient substrate for the UV-induced phosphorylation. Our data also raise the possibility that the 9–1–1 complex plays a negative regulatory role in the Rad17 phosphorylation. We also show that the nucleotide-binding activity of Rad17 is required for its nuclear localization. - Highlights: • We have identified a conserved KYxxL motif in Rad17 protein. • The KYxxL motif is crucial for the interaction with the 9–1–1 complex. • The KYxxL motif is dispensable or inhibitory for UV-induced Rad17 phosphorylation. • Nucleotide binding of Rad17 is required for its nuclear localization.},

  doi          = {10.1016/J.BBRC.2016.07.014},

  url          = {https://www.osti.gov/biblio/22606196},
  journal      = {Biochemical and Biophysical Research Communications},

  issn         = {0006-291X},

  number       = 4,

  volume       = 477,

  place        = {United States},

  year         = {Fri Sep 02 00:00:00 EDT 2016},

  month        = {Fri Sep 02 00:00:00 EDT 2016}

}

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