14-3-3 checkpoint regulatory proteins interact specifically with DNA repair protein human exonuclease 1 (hEXO1) via a semi-conserved motif

Sofie Dabros Andersen, Guido Keijzers, E. Rampakakis, K. Engels, P. Luhn, M. El-shemerly, Finn Cilius Nielsen, YH Du, A. May, V. A. Bohr, S. Ferrari, M. Zannis-Hadjopoulos, H. Fu, Lene Juel Rasmussen

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    Resumé

    Human exonuclease 1 (hEXO1) acts directly in diverse DNA processing events, including replication, mismatch repair (MMR), and double strand break repair (DSBR), and it was also recently described to function as damage sensor and apoptosis inducer following DNA damage. In contrast, 14-3-3 proteins are regulatory phosphorserine/threonine binding proteins involved in the control of diverse cellular events, including cell cycle checkpoint and apoptosis signaling. hEXO1 is regulated by post-translation Ser/Thr phosphorylation in a yet not fully clarified manner, but evidently three phosphorylation sites are specifically induced by replication inhibition leading to protein ubiquitination and degradation. We demonstrate direct and robust interaction between hEXO1 and six of the seven 14-3-3 isoforms in vitro, suggestive of a novel protein interaction network between DNA repair and cell cycle control. Binding experiments reveal weak affinity of the more selective isoform 14-3-3 sigma but both 14-3-3 isoforms eta and sigma significantly stimulate hEXO1 activity, indicating that these regulatory proteins exert a common regulation mode on hEXO1. Results demonstrate that binding involves the phosphorable amino acid S746 in hEXO1 and most likely a second unidentified binding motif. 14-3-3 associations do not appear to directly influence hEXO1 in vitro nuclease activity or in vitro DNA replication initiation. Moreover, specific phosphorylation variants, including hEXO1 S746A, are efficiently imported to the nucleus; to associate with PCNA in distinct replication foci and respond to DNA double strand breaks (DSBs), indicating that 14-3-3 binding does not involve regulating the subcellular distribution of hEXO1. Altogether, these results suggest that association may be related to regulation of hEXO1 availability during the DNA damage response to plausibly prevent extensive DNA resection at the damage site, as supported by recent studies
    OriginalsprogEngelsk
    TidsskriftD N A Repair
    Vol/bind11
    Udgave nummer3
    Sider (fra-til)267-277
    ISSN1568-7864
    DOI
    StatusUdgivet - 2012

    Citer dette

    Andersen, S. D., Keijzers, G., Rampakakis, E., Engels, K., Luhn, P., El-shemerly, M., ... Rasmussen, L. J. (2012). 14-3-3 checkpoint regulatory proteins interact specifically with DNA repair protein human exonuclease 1 (hEXO1) via a semi-conserved motif. D N A Repair, 11(3), 267-277. https://doi.org/10.1016/j.dnarep.2011.11.007
    Andersen, Sofie Dabros ; Keijzers, Guido ; Rampakakis, E. ; Engels, K. ; Luhn, P. ; El-shemerly, M. ; Nielsen, Finn Cilius ; Du, YH ; May, A. ; Bohr, V. A. ; Ferrari, S. ; Zannis-Hadjopoulos, M. ; Fu, H. ; Rasmussen, Lene Juel. / 14-3-3 checkpoint regulatory proteins interact specifically with DNA repair protein human exonuclease 1 (hEXO1) via a semi-conserved motif. I: D N A Repair. 2012 ; Bind 11, Nr. 3. s. 267-277.
    @article{67663c6275ec495e9446072a40584993,
    title = "14-3-3 checkpoint regulatory proteins interact specifically with DNA repair protein human exonuclease 1 (hEXO1) via a semi-conserved motif",
    abstract = "Human exonuclease 1 (hEXO1) acts directly in diverse DNA processing events, including replication, mismatch repair (MMR), and double strand break repair (DSBR), and it was also recently described to function as damage sensor and apoptosis inducer following DNA damage. In contrast, 14-3-3 proteins are regulatory phosphorserine/threonine binding proteins involved in the control of diverse cellular events, including cell cycle checkpoint and apoptosis signaling. hEXO1 is regulated by post-translation Ser/Thr phosphorylation in a yet not fully clarified manner, but evidently three phosphorylation sites are specifically induced by replication inhibition leading to protein ubiquitination and degradation. We demonstrate direct and robust interaction between hEXO1 and six of the seven 14-3-3 isoforms in vitro, suggestive of a novel protein interaction network between DNA repair and cell cycle control. Binding experiments reveal weak affinity of the more selective isoform 14-3-3 sigma but both 14-3-3 isoforms eta and sigma significantly stimulate hEXO1 activity, indicating that these regulatory proteins exert a common regulation mode on hEXO1. Results demonstrate that binding involves the phosphorable amino acid S746 in hEXO1 and most likely a second unidentified binding motif. 14-3-3 associations do not appear to directly influence hEXO1 in vitro nuclease activity or in vitro DNA replication initiation. Moreover, specific phosphorylation variants, including hEXO1 S746A, are efficiently imported to the nucleus; to associate with PCNA in distinct replication foci and respond to DNA double strand breaks (DSBs), indicating that 14-3-3 binding does not involve regulating the subcellular distribution of hEXO1. Altogether, these results suggest that association may be related to regulation of hEXO1 availability during the DNA damage response to plausibly prevent extensive DNA resection at the damage site, as supported by recent studies",
    author = "Andersen, {Sofie Dabros} and Guido Keijzers and E. Rampakakis and K. Engels and P. Luhn and M. El-shemerly and Nielsen, {Finn Cilius} and YH Du and A. May and Bohr, {V. A.} and S. Ferrari and M. Zannis-Hadjopoulos and H. Fu and Rasmussen, {Lene Juel}",
    year = "2012",
    doi = "10.1016/j.dnarep.2011.11.007",
    language = "English",
    volume = "11",
    pages = "267--277",
    journal = "D N A Repair",
    issn = "1568-7864",
    publisher = "Elsevier BV",
    number = "3",

    }

    Andersen, SD, Keijzers, G, Rampakakis, E, Engels, K, Luhn, P, El-shemerly, M, Nielsen, FC, Du, YH, May, A, Bohr, VA, Ferrari, S, Zannis-Hadjopoulos, M, Fu, H & Rasmussen, LJ 2012, '14-3-3 checkpoint regulatory proteins interact specifically with DNA repair protein human exonuclease 1 (hEXO1) via a semi-conserved motif', D N A Repair, bind 11, nr. 3, s. 267-277. https://doi.org/10.1016/j.dnarep.2011.11.007

    14-3-3 checkpoint regulatory proteins interact specifically with DNA repair protein human exonuclease 1 (hEXO1) via a semi-conserved motif. / Andersen, Sofie Dabros; Keijzers, Guido; Rampakakis, E.; Engels, K.; Luhn, P.; El-shemerly, M.; Nielsen, Finn Cilius; Du, YH; May, A.; Bohr, V. A.; Ferrari, S.; Zannis-Hadjopoulos, M.; Fu, H.; Rasmussen, Lene Juel.

    I: D N A Repair, Bind 11, Nr. 3, 2012, s. 267-277.

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    TY - JOUR

    T1 - 14-3-3 checkpoint regulatory proteins interact specifically with DNA repair protein human exonuclease 1 (hEXO1) via a semi-conserved motif

    AU - Andersen, Sofie Dabros

    AU - Keijzers, Guido

    AU - Rampakakis, E.

    AU - Engels, K.

    AU - Luhn, P.

    AU - El-shemerly, M.

    AU - Nielsen, Finn Cilius

    AU - Du, YH

    AU - May, A.

    AU - Bohr, V. A.

    AU - Ferrari, S.

    AU - Zannis-Hadjopoulos, M.

    AU - Fu, H.

    AU - Rasmussen, Lene Juel

    PY - 2012

    Y1 - 2012

    N2 - Human exonuclease 1 (hEXO1) acts directly in diverse DNA processing events, including replication, mismatch repair (MMR), and double strand break repair (DSBR), and it was also recently described to function as damage sensor and apoptosis inducer following DNA damage. In contrast, 14-3-3 proteins are regulatory phosphorserine/threonine binding proteins involved in the control of diverse cellular events, including cell cycle checkpoint and apoptosis signaling. hEXO1 is regulated by post-translation Ser/Thr phosphorylation in a yet not fully clarified manner, but evidently three phosphorylation sites are specifically induced by replication inhibition leading to protein ubiquitination and degradation. We demonstrate direct and robust interaction between hEXO1 and six of the seven 14-3-3 isoforms in vitro, suggestive of a novel protein interaction network between DNA repair and cell cycle control. Binding experiments reveal weak affinity of the more selective isoform 14-3-3 sigma but both 14-3-3 isoforms eta and sigma significantly stimulate hEXO1 activity, indicating that these regulatory proteins exert a common regulation mode on hEXO1. Results demonstrate that binding involves the phosphorable amino acid S746 in hEXO1 and most likely a second unidentified binding motif. 14-3-3 associations do not appear to directly influence hEXO1 in vitro nuclease activity or in vitro DNA replication initiation. Moreover, specific phosphorylation variants, including hEXO1 S746A, are efficiently imported to the nucleus; to associate with PCNA in distinct replication foci and respond to DNA double strand breaks (DSBs), indicating that 14-3-3 binding does not involve regulating the subcellular distribution of hEXO1. Altogether, these results suggest that association may be related to regulation of hEXO1 availability during the DNA damage response to plausibly prevent extensive DNA resection at the damage site, as supported by recent studies

    AB - Human exonuclease 1 (hEXO1) acts directly in diverse DNA processing events, including replication, mismatch repair (MMR), and double strand break repair (DSBR), and it was also recently described to function as damage sensor and apoptosis inducer following DNA damage. In contrast, 14-3-3 proteins are regulatory phosphorserine/threonine binding proteins involved in the control of diverse cellular events, including cell cycle checkpoint and apoptosis signaling. hEXO1 is regulated by post-translation Ser/Thr phosphorylation in a yet not fully clarified manner, but evidently three phosphorylation sites are specifically induced by replication inhibition leading to protein ubiquitination and degradation. We demonstrate direct and robust interaction between hEXO1 and six of the seven 14-3-3 isoforms in vitro, suggestive of a novel protein interaction network between DNA repair and cell cycle control. Binding experiments reveal weak affinity of the more selective isoform 14-3-3 sigma but both 14-3-3 isoforms eta and sigma significantly stimulate hEXO1 activity, indicating that these regulatory proteins exert a common regulation mode on hEXO1. Results demonstrate that binding involves the phosphorable amino acid S746 in hEXO1 and most likely a second unidentified binding motif. 14-3-3 associations do not appear to directly influence hEXO1 in vitro nuclease activity or in vitro DNA replication initiation. Moreover, specific phosphorylation variants, including hEXO1 S746A, are efficiently imported to the nucleus; to associate with PCNA in distinct replication foci and respond to DNA double strand breaks (DSBs), indicating that 14-3-3 binding does not involve regulating the subcellular distribution of hEXO1. Altogether, these results suggest that association may be related to regulation of hEXO1 availability during the DNA damage response to plausibly prevent extensive DNA resection at the damage site, as supported by recent studies

    U2 - 10.1016/j.dnarep.2011.11.007

    DO - 10.1016/j.dnarep.2011.11.007

    M3 - Journal article

    VL - 11

    SP - 267

    EP - 277

    JO - D N A Repair

    JF - D N A Repair

    SN - 1568-7864

    IS - 3

    ER -