Active role for nibrin in the kinetics of Atm activation

被引:65
作者
Cerosaletti, K
Wright, J
Concannon, P
机构
[1] Benaroya Res Inst, Mol Genet Program, Seattle, WA 98101 USA
[2] Univ Washington, Sch Med, Dept Immunol, Seattle, WA 98195 USA
关键词
D O I
10.1128/MCB.26.5.1691-1699.2006
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The Atm protein kinase is central to the DNA double-strand break response in mammalian cells. After irradiation, dimeric Atm undergoes autophosphorylation at Ser 1981 and dissociates into active monomers. Atm activation is stimulated by expression of the Mre11/Rad50/nibrin complex. Previously, we showed that a C-terminal fragment of nibrin, containing binding sites for both Well and Atm, was sufficient to provide this stimulatory effect in Nijmegen breakage syndrome (NBS) cells. To discriminate whether nibrin's role in Atm activation is to bind and translocate Mre11/Rad50 to the nucleus or to interact directly with Atm, we expressed an Mre11 transgene with a C-terminal NLS sequence in NBS fibroblasts. The Mre11-NLS protein complexed with Rad50, localized to the nucleus in NBS fibroblasts, and associated with chromatin. However, Atm autophosphorylation was not stimulated in cells expressing Mre11-NLS, nor were downstream Atm targets phosphorylated. To determine whether nibrin-Atm interaction is necessary to stimulate Atm activation, we expressed nibrin transgenes lacking the Atm binding domain in NBS fibroblasts. The nibrin Delta Atm protein interacted with Mre11/Rad50; however, Atm autophosphorylation was dramatically reduced after irradiation in NBS cells expressing the nibrin Delta Atm transgenes relative to wild-type nibrin. These results indicate that nibrin plays an active role in Atm activation beyond translocating Mre11/Rad50 to the nucleus and that this function requires nibrin-Atm interaction.
引用
收藏
页码:1691 / 1699
页数:9
相关论文
共 46 条
[21]   Immortalization and characterization of Nijmegen Breakage Syndrome fibroblasts [J].
Kraakman-van der Zwet, M ;
Overkamp, WJI ;
Friedl, AA ;
Klein, B ;
Verhaegh, GWCT ;
Jaspers, NGJ ;
Midro, AT ;
Eckardt-Schupp, F ;
Lohman, PHM ;
Zdzienicka, MZ .
MUTATION RESEARCH-DNA REPAIR, 1999, 434 (01) :17-27
[22]   Regulation of Mre11/Rad50 by Nbs1 - Effects on nucleotide-dependent DNA binding and association with ataxia-telangiectasia-like disorder mutant complexes [J].
Lee, JH ;
Ghirlando, R ;
Bhaskara, V ;
Hoffmeyer, MR ;
Gu, J ;
Paull, TT .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (46) :45171-45181
[23]   Direct activation of the ATM protein kinase by the Mre11/Rad50/Nbs1 complex [J].
Lee, JH ;
Paull, TT .
SCIENCE, 2004, 304 (5667) :93-96
[24]   ATM activation by DNA double-strand breaks through the Mre11-Rad50-Nbs1 complex [J].
Lee, JH ;
Paull, TT .
SCIENCE, 2005, 308 (5721) :551-554
[25]   ATM phosphorylates p95/nbs1 in an S-phase checkpoint pathway [J].
Lim, DS ;
Kim, ST ;
Xu, B ;
Maser, RS ;
Lin, JY ;
Petrini, JHJ ;
Kastan, MB .
NATURE, 2000, 404 (6778) :613-+
[26]   Distinct spatiotemporal dynamics of mammalian checkpoint regulators induced by DNA damage [J].
Lukas, C ;
Falck, J ;
Bartkova, J ;
Bartek, J ;
Lukas, J .
NATURE CELL BIOLOGY, 2003, 5 (03) :255-U12
[27]   Mdc1 couples DNA double-strand break recognition by Nbs1 with its H2AX-dependent chromatin retention [J].
Lukas, C ;
Melander, F ;
Stucki, M ;
Falck, J ;
Bekker-Jensen, S ;
Goldberg, M ;
Lerenthal, Y ;
Jackson, SP ;
Bartek, J ;
Lukas, J .
EMBO JOURNAL, 2004, 23 (13) :2674-2683
[28]   An alternative mode of translation permits production of a variant NBS1 protein from the common Nijmegen breakage syndrome allele [J].
Maser, RS ;
Zinkel, R ;
Petrini, JHJ .
NATURE GENETICS, 2001, 27 (04) :417-421
[29]   Ataxia telangiectasia-mutated phosphorylates Chk2 in vivo and in vitro [J].
Matsuoka, S ;
Rotman, G ;
Ogawa, A ;
Shiloh, Y ;
Tamai, K ;
Elledge, SJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (19) :10389-10394
[30]   Chromatin association of human origin recognition complex, Cdc6, and minichromosome maintenance proteins during the cell cycle:: Assembly of prereplication complexes in late mitosis [J].
Méndez, J ;
Stillman, B .
MOLECULAR AND CELLULAR BIOLOGY, 2000, 20 (22) :8602-8612