Functional characterization of Rad18 domains for Rad6, ubiquitin, DNA binding and PCNA modification

Research output: Contribution to journal › Journal article › Research › peer-review

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Functional characterization of Rad18 domains for Rad6, ubiquitin, DNA binding and PCNA modification. / Notenboom, Valerie; Hibbert, Richard G; van Rossum-Fikkert, Sarah E; Olsen, Jesper Velgaard; Mann, Matthias; Sixma, Titia K.

In: Nucleic Acids Research, Vol. 35, No. 17, 2007, p. 5819-30.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Notenboom, V, Hibbert, RG, van Rossum-Fikkert, SE, Olsen, JV, Mann, M & Sixma, TK 2007, 'Functional characterization of Rad18 domains for Rad6, ubiquitin, DNA binding and PCNA modification', Nucleic Acids Research, vol. 35, no. 17, pp. 5819-30. https://doi.org/10.1093/nar/gkm615

APA

Notenboom, V., Hibbert, R. G., van Rossum-Fikkert, S. E., Olsen, J. V., Mann, M., & Sixma, T. K. (2007). Functional characterization of Rad18 domains for Rad6, ubiquitin, DNA binding and PCNA modification. Nucleic Acids Research, 35(17), 5819-30. https://doi.org/10.1093/nar/gkm615

Vancouver

Notenboom V, Hibbert RG, van Rossum-Fikkert SE, Olsen JV, Mann M, Sixma TK. Functional characterization of Rad18 domains for Rad6, ubiquitin, DNA binding and PCNA modification. Nucleic Acids Research. 2007;35(17):5819-30. https://doi.org/10.1093/nar/gkm615

Author

Notenboom, Valerie ; Hibbert, Richard G ; van Rossum-Fikkert, Sarah E ; Olsen, Jesper Velgaard ; Mann, Matthias ; Sixma, Titia K. / Functional characterization of Rad18 domains for Rad6, ubiquitin, DNA binding and PCNA modification. In: Nucleic Acids Research. 2007 ; Vol. 35, No. 17. pp. 5819-30.

Bibtex

@article{9d11bfd583aa413a82c71cd394872b82,
title = "Functional characterization of Rad18 domains for Rad6, ubiquitin, DNA binding and PCNA modification",
abstract = "Rad18 is a ubiquitin E3 ligase that monoubiquitinates PCNA on stalled replications forks. This allows recruitment of damage-tolerant polymerases for damage bypass and DNA repair. In this activity, the Rad18 protein has to interact with Rad6, the E2 ubiquitin-conjugating enzyme, ubiquitin, PCNA and DNA. Here we analyze the biochemical interactions of specific domains of the Rad18 protein. We found that the Rad6/Rad18 complex forms stable dimers in vitro. Consistent with previous findings, both the Ring domain and a C-terminal region contribute to the Rad6 interaction, while the C-terminus is not required for the interaction with PCNA. Surprisingly we find that the C2HC zinc finger is important for interaction with ubiquitin, apparently analogous to the interactions of classical zinc fingers with ubiquitin such as found in the UBZ and UBM domains in Y-family polymerases. Finally we find that the SAP domain, but not the zinc finger domain, is capable of DNA binding in vitro.",
author = "Valerie Notenboom and Hibbert, {Richard G} and {van Rossum-Fikkert}, {Sarah E} and Olsen, {Jesper Velgaard} and Matthias Mann and Sixma, {Titia K}",
note = "Keywords: Animals; Binding Sites; DNA; DNA-Binding Proteins; Dimerization; Mice; Proliferating Cell Nuclear Antigen; Protein Structure, Tertiary; Ubiquitin; Ubiquitin-Conjugating Enzymes",
year = "2007",
doi = "10.1093/nar/gkm615",
language = "English",
volume = "35",
pages = "5819--30",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "17",

}

RIS

TY - JOUR

T1 - Functional characterization of Rad18 domains for Rad6, ubiquitin, DNA binding and PCNA modification

AU - Notenboom, Valerie

AU - Hibbert, Richard G

AU - van Rossum-Fikkert, Sarah E

AU - Olsen, Jesper Velgaard

AU - Mann, Matthias

AU - Sixma, Titia K

N1 - Keywords: Animals; Binding Sites; DNA; DNA-Binding Proteins; Dimerization; Mice; Proliferating Cell Nuclear Antigen; Protein Structure, Tertiary; Ubiquitin; Ubiquitin-Conjugating Enzymes

PY - 2007

Y1 - 2007

N2 - Rad18 is a ubiquitin E3 ligase that monoubiquitinates PCNA on stalled replications forks. This allows recruitment of damage-tolerant polymerases for damage bypass and DNA repair. In this activity, the Rad18 protein has to interact with Rad6, the E2 ubiquitin-conjugating enzyme, ubiquitin, PCNA and DNA. Here we analyze the biochemical interactions of specific domains of the Rad18 protein. We found that the Rad6/Rad18 complex forms stable dimers in vitro. Consistent with previous findings, both the Ring domain and a C-terminal region contribute to the Rad6 interaction, while the C-terminus is not required for the interaction with PCNA. Surprisingly we find that the C2HC zinc finger is important for interaction with ubiquitin, apparently analogous to the interactions of classical zinc fingers with ubiquitin such as found in the UBZ and UBM domains in Y-family polymerases. Finally we find that the SAP domain, but not the zinc finger domain, is capable of DNA binding in vitro.

AB - Rad18 is a ubiquitin E3 ligase that monoubiquitinates PCNA on stalled replications forks. This allows recruitment of damage-tolerant polymerases for damage bypass and DNA repair. In this activity, the Rad18 protein has to interact with Rad6, the E2 ubiquitin-conjugating enzyme, ubiquitin, PCNA and DNA. Here we analyze the biochemical interactions of specific domains of the Rad18 protein. We found that the Rad6/Rad18 complex forms stable dimers in vitro. Consistent with previous findings, both the Ring domain and a C-terminal region contribute to the Rad6 interaction, while the C-terminus is not required for the interaction with PCNA. Surprisingly we find that the C2HC zinc finger is important for interaction with ubiquitin, apparently analogous to the interactions of classical zinc fingers with ubiquitin such as found in the UBZ and UBM domains in Y-family polymerases. Finally we find that the SAP domain, but not the zinc finger domain, is capable of DNA binding in vitro.

U2 - 10.1093/nar/gkm615

DO - 10.1093/nar/gkm615

M3 - Journal article

VL - 35

SP - 5819

EP - 5830

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 17

ER -

ID: 46461130