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1. Edited by George Klein, Karolinska Institute, Stockholm, Sweden, and approved December 8, 2009 (received
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Abstract
Ataxia-telangiectasia mutated (ATM) is a high molecular weight protein serine/threonine kinase
that plays a central role in the maintenance of genomic integrity by activating cell cycle
checkpoints and promoting repair of DNA double-strand breaks. Little is known about the
regulatory mechanisms for ATM expression itself. MicroRNAs are naturally existing regulators
that modulate gene expression in a sequence-specific manner. Here, we show that a human
microRNA, miR-421, suppresses ATM expression by targeting the 3′-untranslated region (3′UTR)
of ATM transcripts. Ectopic expression of miR-421 resulted in S-phase cell cycle checkpoint
changes and an increased sensitivity to ionizing radiation, creating a cellular phenotype similar to
that of cells derived from ataxia-telangiectasia (A-T) patients. Blocking the interaction between
miR-421 and ATM 3′UTR with an antisense morpholino oligonucleotide rescued the defective
phenotype caused by miR-421 overexpression, indicating that ATM mediates the effect of miR-
421 on cell cycle checkpoint and radiosensitivity. Overexpression of the N-Myc transcription
factor, an oncogene frequently amplified in neuroblastoma, induced miR-421 expression, which,
in turn, down-regulated ATM expression, establishing a linear signaling pathway that may
contribute to N-Myc-induced tumorigenesis in neuroblastoma. Taken together, our findings
implicate a previously undescribed regulatory mechanism for ATM expression and ATM-
dependent DNA damage response and provide several potential targets for treating
neuroblastoma and perhaps A-T.
• neuroblastoma
• S-phase checkpoint
• radiosensitivity
• DNA repair
Ataxia-telangiectasia mutated (ATM) kinase plays a hierarchical regulatory role in the double-
strand break (DSB)-induced DNA damage response in which ATM transduces a DSB
damage/repair signal to downstream effector machinery by phosphorylating critical protein
substrates (1 –4). ATM mutations, which usually result in loss of ATM protein expression (5), lead
to the autosomal recessive progressive neurodegenerative disease ataxia-telangiectasia (A-T) (6,
7). Both homozygotes and heterozygotes are at an increased risk for cancer (8). ATM has been
reported to be regulated by a transcription factor, E2F-1, (9) and the ATM gene is also reported to
be subject to epigenetic silencing such as by methylation of the ATM promoter (10, 11),
suggesting that ATM can also be up-regulated at the transcriptional level under some
circumstances. MicroRNAs regulate gene expression through inhibition of translation or
degradation of the targeted mRNA (12, 13). Physiological functions of microRNAs have been
observed in normal and lineage-targeted development (14) as well as in the context of human
cancers (15). In this study, we demonstrate that miR-421 targets the 3′-untranslated region
(3′UTR) of ATM and down-regulates its expression, whereas miR-421 expression is driven by the
N-Myc transcription factor, an oncogene that is frequently amplified in neuroblastoma cells.
• In a new window
Fig. 1.
miR-421 suppresses ATM expression by targeting ATM 3′UTR. (A) Mature miR-421 sequences and
recognition sites within 3′UTR of ATM. The seed sequence of miR-421 is shown in the box. WT and
del6 (Δ6) ATM-3′UTR targets are also shown. (B) Constructs of Renilla luciferase [Luc; unmodified
construct (pRL)-CMV] containing WT or 6-nt deleted (Δ6) ATM 3′UTR. (C) Luciferase (Luc.) activity
of pRL and modified constructs containing WT or mutant (Δ6) 3′UTR. Luciferase constructs were
cotransfected with pre-miR-CTL (control, 50 nM) or pre-miR-421 (50 nM) into HeLa cells. Renilla
luciferase activity was measured 36 h after incubation and normalized to firefly luciferase.
Asterisk indicates significant down-regulation of pre-miR-421 against construct containing WT
ATM 3′UTR. (D) Immunoblot of endogenous ATM expression in HeLa cells 96 h after transfection
of increasing amounts of pre-miR-421 (using pre-miR-CTL to compensate for equal amounts of
total miRs). SMC1 served as a loading control for the blot. (E) Immunoblot of pS966-SMC1 in HeLa
cells that were transiently transfected with pre-miR-CTL (100 nM) or pre-miR-421 (100 nM) after
10-Gy IR to activate the DNA damage response. A WT lymphoblastoid cell line (LCL) served as a
positive control for SMC1 and ATM protein. (F) Real-time PCR of ATM mRNA from HeLa cells
transfected with pre-miR-CTL (100 nM) or pre-miR-421 (100n M). Data were normalized to the
level of GAPDH mRNA, and the ratio of ATM/GAPDH in HeLa control cells was set to 1.
• In a new window
Fig. 2.
miR-421 regulates cell cycle S-phase checkpoint and cellular radiosensitivity. (A) Scheme of a U6
promoter-driven miR-421 cloned into a lentiviral vector with two LTRs and a selection marker for
blasticidin driven by SV40 promoter. (B) Real-time PCR of miR-421 expression in HeLa cells stably
overexpressing scrambled shRNA (HeLa/scram) or miR-421(HeLa/miR-421). Data were
normalized to an internal control RNU66, and the ratio of miR-421/RNU66 in HeLa/scram cells was
set to 1. (C) Immunoblot of ATM and pSMC1 in HeLa/scram and HeLa/miR-421 cells with or
without 10-Gy IR. Note the reduction of both ATM and pSMC1 in miR421-overexpressing cells. (D)
Analysis of IR-induced cell cycle S-phase checkpoint by FC. Stably overexpressing HeLa/scram
and HeLa/miR-421 cells were treated with or without 10 Gy. DNA synthesis at S-phase was
labeled with BrdU. (Left Upper and Lower) Results of one experiment representative of three
independent experiments. Box R5 indicates the percentage of BrdU+ S-phase cells pre- or post-IR.
(Right) Summary of change of BrdU+ cells pre- and post-IR for HeLa/scram and HeLa/miR-421
cells from three independent experiments, using the algorithm (R5+IR−R5−IR)/R5−IR × 100%. (E)
HeLa/scram and HeLa/miR-421 cells were irradiated at the indicated doses, and colony survival
was measured after 2 weeks. (F) Effect of miR-421 on proliferation of HeLa cells, as measured by
cell population doublings with culture time. (G) Effect of miR-421 on IR-induced cell death, as
measured by propidium iodide staining FC. The percentage of propidium iodide-positive cells was
normalized to the unirradiated cells in each group.
ATM regulates DNA damage-induced cell cycle checkpoints at G1-S and intra-S phase (18, 19). A
hallmark of A-T cells is the failure to arrest DNA synthesis in S-phase following DNA damage and
continuous incorporation of nucleotides into DNA despite damage (i.e., radioresistant DNA
synthesis) (20, 21). Thus, we anticipated that miR-421 overexpression might regulate DNA
damage-induced cell cycle S-phase checkpoints. To assess this, HeLa/scram and HeLa/miR-421
cells were irradiated (10 Gy) to introduce DNA damage and BrdU was used to follow DNA
incorporation. As expected, a reduction in the percentage of BrdU-positive cells in S-phase was
observed for the HeLa/scram control cells (14.88% pre-IR vs. 12.56% post-IR), indicating a normal
block in the DNA synthesis (Fig. 2D , Upper Left and Right); in contrast, an increase in the
percentage of BrdU-positive cells in S-phase was observed for HeLa/miR-421 cells (11.67% pre-IR
vs. 15.38% post-IR) (Fig. 2D , Lower Left and Right), indicating that miR-421 overexpression
overcomes the IR-induced DNA synthesis block and mimics the radioresistant DNA synthesis of A-
T cells. The miR421-induced continuous DNA synthesis was also seen with lower doses of IR at 2
and 5 Gy (Fig. S1A ). We noticed that the pre-IR percentage of HeLa/miR-421 cells in S-phase was
lower than that of control HeLa/scram cells, suggesting that miR-421 might regulate this cell
cycle checkpoint independent of DNA damage. Similar results were observed using a human
breast cancer cell line, MDA-MB-231, when miR-421 was overexpressed (Fig. S2 A and B ).
A clonogenic assay was used to determine whether overexpression of miR-421 affects cellular
radiosensitivity. As shown in Fig. 2E , the survival fractions of HeLa/miR-421 cells post-IR (1 and 2
Gy) were significantly reduced relative to those of HeLa/scram control cells. MiR-421
overexpression did not alter the proliferation rate of HeLa cells (Fig. 2F ) but increased post-IR
cell death (Fig. 2G ), which is consistent with the decreased survival fraction in the clonogenic
assay. A similar effect of miR-421 on radiosensitivity was observed with MDA-MB-231 cells (Fig.
S2C ).
• In a new window
Fig. 3.
ATM mediates the effect of miR-421 on cell cycle S-phase checkpoint and radiosensitivity. (A)
Schematic working model of ATM 3′UTR that was targeted by an antisense AMO. AMO-ATM was
designed to match the miR-421 recognition site of ATM 3′UTR and specifically block the down-
regulation of ATM by impeding the binding of mature miR-421. (B) (Left) Immunoblot of ATM
expression in HeLa/scram and HeLa/miR-421 cells treated with or without AMO-ATM (2 μM) for 5
days. The fold change in ATM expression is shown below the immunoblot. (Right) ELISA was also
used to determine ATM concentration. (C) Immunoblot of pSMC1 in HeLa/scram and HeLa/miR-
421 cells treated with AMO-scram (2 μM) or AMO-ATM (2 μM) for 5 days, followed by 10-Gy IR.
The fold change in pSMC1 level is shown below the immunoblot. Note the increase of pSMC1 in
HeLa/miR-421 cells treated with AMO-ATM. (D) Analysis of cell cycle S-phase checkpoint after
treatment of AMO. HeLa/scram and HeLa/miR-421 cells were treated with AMO-scram (2 μM) or
AMO-ATM (2 μM) for 5 days and irradiated with increasing doses of radiation (2, 5, and 10 Gy).
DNA synthesis was monitored by BrdU incorporation and analyzed by FC. The percentage of
BrdU+ S-phase cells at the start point (unirradiated) was arbitrarily set to 50%, and all other data
were normalized to this point. This plot is representative of three independent experiments. The
arrow indicates that AMO-ATM treatment rescues the defect of HeLa/miR-421 cells. (E) Colony
survival fraction with exposure to AMO. HeLa/scram and HeLa/miR-421 cells were treated with
AMO-scram (2 μM) or AMO-ATM (2 μM) for 5 days, and 500 cells were plated in triplicate; cells
were irradiated with increasing doses of radiation, and surviving colonies were scored after 2
weeks. The survival fraction at each radiation dose was normalized to that of the nonirradiated
control. The arrow indicates that the AMO rescued the radiosensitivity of HeLa/miR-421 cells.
Fig. 4.
miR-421 is up-regulated by N-Myc overexpression in HeLa cells. (A) Chromosomal location of miR-
374b/miR-421 cluster on chromosome Xq13, sharing the same promoter. The promoter region (1
kb), containing an E-box (5′-CACGTG-3′), was cloned into luciferase construct pGL3-basic to
create pGL3-PR421 and drives the transcription of firefly luciferase (Luc). (B) Luciferase activity of
the miR-421 promoter. Luciferase constructs [pGL3-PR421 and unmodified construct (pRL)-CMV]
were cotransfected, with vector (Vec) or N-Myc, into HeLa cells. Firefly luciferase activity was
measured 24 h and 48 h after incubation and normalized to Renilla luciferase activity. (C) Real-
time PCR of endogenous miR-421expression in HeLa cells transiently transfected with vector or
N-Myc. Data were normalized to RNU66. (D) Immunoblot of ATM expression in HeLa cells
transiently transfected with increasing amounts of N-Myc (using vector to compensate for equal
amounts of total DNA). The fold change in ATM protein expression is shown below the blot. (E)
ELISA measurement of ATM concentrations in HeLa cells transiently transfected with N-Myc. The
asterisk indicates significant inhibition of ATM by N-Myc overexpression. (F) ELISA to determine
ATM concentration in HeLa cells transiently transfected with the indicated DNA constructs (vector
or N-Myc) and anti-miR-CTL (50 nM) or anti-miR-421(50 nM) inhibitors. (G) Immunoblot of ATM
expression in HeLa cells transiently transfected with indicated DNA constructs (vector or N-Myc)
and anti-miR-CTL (50 nM) or anti-miR-421(50 nM) inhibitor. Only the top band corresponds to
ATM.
• In a new window
Fig. 5.
N-Myc negatively regulates ATM via miR-421 in neuroblastoma cells. (A) Immunoblot of ATM
expression in N-Myc-amplified (CHLA-134, CHLA136, LA-N-5, and LA-N-1) or -nonamplified (CHLA-
15, CHLA-90, and CHLA-255) neuroblastoma cell lines. We observed some N-Myc expression in
CHLA-90, although it is an N-Myc-nonamplified cell line; ATM expression was relatively lower in
this cell line when compared with CHLA-15 or CHLA-255. A-T lymphoblastoid cells (AT-LCL) and
WT lymphoblastoid cells (WT-LCL) are negative and positive controls, respectively, for ATM
expression. In total, 100 μg of total protein for all neuroblastoma cells and only 25 μg of total
protein for AT-LCL and WT-LCL were loaded; SMC1 served as a loading control. (B) ChIP PCR
assay detects the in vivo binding of N-Myc protein to the miR-421 promoter DNA. A PCR fragment
of expected size (246 bp) was seen in the N-Myc-amplified (amp.) LA-N-1 cells
immunoprecipitated with the specific anti-N-Myc antibody (lane 5) but not without antibody or
with nonspecific mouse IgG (lanes 2 and 3). No signal was seen in the N-Myc-nonamplified CHLA-
255 cells immunoprecipitated with no antibody, nonspecific mouse IgG, or specific anti-Myc
antibody (lanes 7–9). PCR with input DNA was used as a positive control. (C) Real-time PCR of
endogenous miR-421 in the N-Myc-amplified LA-N-1 cells and the N-Myc-nonamplified CHLA-255
cells. RNU66 was used as an internal control. (D) Immunoblot of ATM expression in CHLA-255 and
LA-N-1 cells treated with AMO-scram (4 μM) or AMO-ATM (4 μM) for 5 days or in L-AN-1 cells
transfected with anti-miR-CTL (100 nM) or anti-miR-421 inhibitor (100 nM) for 96 h. The fold
change in ATM expression is shown below. (E) Real-time PCR of miR-421 expression in LA-N-1
cells treated with AMO-scram (4 μM) or AMO-ATM (4 μM) for 5 days or transfected with anti-miR-
CTL (100 nM) or anti-miR-421 (100 nM) inhibitor for 4 days. RNU66 was used as an internal
control. (F) FC-SMC1 detection of IR-induced ATM-dependent phosphorylation of SMC1 in AMO-
treated neuroblastoma cells. LA-N-1 and CHLA-255 cells were treated with AMO-scram (4 μM) or
AMO-ATM (4 μM) for 5 days and subjected to 10-Gy IR. The pSMC1 level is indicated by the
fluorescence intensity. The filled peaks represent the cells without IR, and unfilled peaks
represent post-IR cells. This panel is representative of three independent experiments. (G) Linear
signaling pathway in which N-Myc up-regulates miR-421 expression and miR-421, in turn, down-
regulates ATM expression by targeting its 3′UTR.
Because c-Myc shares a conserved E-box binding site (5′-CACGTG-3′) with N-Myc (Fig. 4A ), we
were prompted to determine whether c-Myc functions in a manner similar to N-Myc in up-
regulating miR-421 expression. As shown in Fig. S4A , cotransfection of c-Myc with the miR-421
promoter construct into HeLa cells resulted in a significant increase in miR-421 promoter-driven
luciferase activity, as did cotransfection of N-Myc. Endogenous miR-421 expression in HeLa cells
was similarly increased ∼1.5-fold after transfection of c-Myc (Fig. S4B ).
We have established that miR-421 expression is up-regulated by the transcription factor N-Myc,
establishing a linear signaling pathway (N-Myc → miR-421 → ATM) in such a manner that the
oncogene N-Myc negatively regulates the tumor suppressor ATM (Fig. 5G ). Because the ATM-
driven DNA damage response is thought to be a physiological barrier in early human
tumorigenesis (30 –33), our findings add that miR421-mediated ATM down-regulation may
contribute to N-Myc-induced tumorigenesis in neuroblastoma. The finding that the up-regulation
of miR-421 can alter the cellular radiosensitivity suggests that treatment of proliferating cancer
cells with miR-421-inducing agents might sensitize them for radiotherapy. Conversely the finding
that exposure of neuroblastoma cells to AMO-ATM increases ATM expression implies that AMO-
ATM holds therapeutical potential for N-Myc-amplified neuroblastomas, perhaps by enhancing
ATM-dependent apoptosis in response to DNA damage (34, 35) or driving nondividing
differentiated neuronal cells to reenter S-phase (36). Lastly, the suppression of ATM by miR-421
introduces two possible pathogenetic mechanisms for A-T: A mutation in the ATM 3′UTR might
enhance the binding of miR-421, or a mutation of miR-421 might result in miR-421
overexpression, both leading to the down-regulation of ATM expression. Such disease-causing
mutations of microRNA-binding sites in the 3′UTR of the target genes have been reported (37).
However, no such mutations have been observed to date in A-T patients. Our findings also
suggest that miR-421 could function as a modifier gene, contributing to the A-T phenotype and
perhaps to the variability of disease onset and progression.
The transcription factor binding site analysis was done using the CONSITE database. A 2-kb
genomic sequence upstream of miR-421 was used as the analyzing template, and the cutoff
value for transcription factors was set to 99%; N-Myc transcription factor was the top candidate.
ATM-ELISA.
An ELISA was used to determine the relative ATM expression in cells and performed as previously
described (39). More details on the ATM-ELISA are provided in SI Text .
• Author contributions: H.H. and R.A.G. designed research; H.H., L.D., and G.N.
performed research; R.C.S. contributed new reagents/analytic tools; H.H. and
R.A.G. analyzed data; and H.H., R.C.S., and R.A.G. wrote the paper.
Previous Section
References
1. ↵
1. Shiloh Y
(2003) ATM and related protein kinases: Safeguarding genome integrity. Nat Rev
Cancer 3:155–168.
CrossRefMedlineWeb of Science
2. ↵
1. Lavin MF
(2008) Ataxia-telangiectasia: From a rare disorder to a paradigm for cell signalling
and cancer. Nat Rev Mol Cell Biol 9:759–769.
CrossRefMedlineWeb of Science
3. ↵
1. Matsuoka S,
2. et al.
(2007) ATM and ATR substrate analysis reveals extensive protein networks
responsive to DNA damage. Science 316:1160–1166.
Abstract/FREE Full Text
4. ↵
1. Bakkenist CJ,
2. Kastan MB
(2003) DNA damage activates ATM through intermolecular autophosphorylation and
dimer dissociation. Nature 421:499–506.
CrossRefMedline
5. ↵
1. Concannon P,
2. Gatti RA
(1997) Diversity of ATM gene mutations detected in patients with ataxia-
telangiectasia. Hum Mutat 10:100–107.
CrossRefMedlineWeb of Science
6. ↵
1. Gatti RA,
2. et al.
(1988) Localization of an ataxia-telangiectasia gene to chromosome 11q22-23.
Nature 336:577–580.
CrossRefMedline
7. ↵
1. Savitsky K,
2. et al.
(1995) A single ataxia telangiectasia gene with a product similar to PI-3 kinase.
Science 268:1749–1753.
Abstract/FREE Full Text
8. ↵
1. Swift M,
2. Reitnauer PJ,
3. Morrell D,
4. Chase CL
(1987) Breast and other cancers in families with ataxia-telangiectasia. N Engl J Med
316:1289–1294.
MedlineWeb of Science
9. ↵
1. Berkovich E,
2. Ginsberg D
(2003) ATM is a target for positive regulation by E2F-1. Oncogene 22:161–167.
CrossRefMedlineWeb of Science
10. ↵
1. Roy K,
2. Wang L,
3. Makrigiorgos GM,
4. Price BD
(2006) Methylation of the ATM promoter in glioma cells alters ionizing radiation
sensitivity. Biochem Biophys Res Commun 344:821–826.
CrossRefMedlineWeb of Science
11. ↵
1. Kim WJ,
2. Vo QN,
3. Shrivastav M,
4. Lataxes TA,
5. Brown KD
(2002) Aberrant methylation of the ATM promoter correlates with increased
radiosensitivity in a human colorectal tumor cell line. Oncogene 21:3864–3871.
CrossRefMedlineWeb of Science
12. ↵
1. He L,
2. Hannon GJ
(2004) MicroRNAs: Small RNAs with a big role in gene regulation. Nat Rev Genet
5:522–531.
CrossRefMedline
13. ↵
1. Bartel DP
(2009) MicroRNAs: Target recognition and regulatory functions. Cell 136:215–233.
CrossRefMedlineWeb of Science
14. ↵
1. Stefani G,
2. Slack FJ
(2008) Small non-coding RNAs in animal development. Nat Rev Mol Cell Biol 9:219–
230.
CrossRefMedlineWeb of Science
15. ↵
1. Calin GA,
2. Croce CM
(2006) MicroRNA-cancer connection: The beginning of a new tale. Cancer Res
66:7390–7394.
Abstract/FREE Full Text
16. ↵
1. Kitagawa R,
2. Bakkenist CJ,
3. McKinnon PJ,
4. Kastan MB
(2004) Phosphorylation of SMC1 is a critical downstream event in the ATM-NBS1-
BRCA1 pathway. Genes Dev 18:1423–1438.
Abstract/FREE Full Text
17. ↵
1. Hu H,
2. et al.
(2008) Integration of transforming growth factor beta and RAS signaling silences a
RAB5 guanine nucleotide exchange factor and enhances growth factor-directed cell
migration. Mol Cell Biol 28:1573–1583.
Abstract/FREE Full Text
18. ↵
1. Zhou BB,
2. Elledge SJ
(2000) The DNA damage response: Putting checkpoints in perspective. Nature
408:433–439.
CrossRefMedline
19. ↵
1. Kastan MB,
2. Bartek J
(2004) Cell-cycle checkpoints and cancer. Nature 432:316–323.
CrossRefMedline
20. ↵
1. Houldsworth J,
2. Lavin MF
(1980) Effect of ionizing radiation on DNA synthesis in ataxia telangiectasia cells.
Nucleic Acids Res 8:3709–3720.
Abstract/FREE Full Text
21. ↵
1. Painter RB
(1981) Radioresistant DNA synthesis: An intrinsic feature of ataxia telangiectasia.
Mutat Res 84:183–190.
CrossRefMedlineWeb of Science
22. ↵
1. Krek A,
2. et al.
(2005) Combinatorial microRNA target predictions. Nat Genet 37:495–500.
CrossRefMedlineWeb of Science
23. ↵
1. Seeger RC,
2. et al.
(1985) Association of multiple copies of the N-myc oncogene with rapid progression
of neuroblastomas. N Engl J Med 313:1111–1116.
MedlineWeb of Science
24. ↵
1. Brodeur GM
(2003) Neuroblastoma: Biological insights into a clinical enigma. Nat Rev Cancer
3:203–216.
CrossRefMedlineWeb of Science
25. ↵
1. Nahas SA,
2. Butch AW,
3. Du L,
4. Gatti RA
(2009) Rapid flow cytometry-based structural maintenance of chromosomes 1
(SMC1) phosphorylation assay for identification of ataxia-telangiectasia
homozygotes and heterozygotes. Clin Chem 55:463–472.
Abstract/FREE Full Text
26. ↵
1. Basso K,
2. et al.
(2009) Identification of the human mature B cell miRNome. Immunity 30:744–752.
CrossRefMedlineWeb of Science
27. ↵
1. Klein U,
2. Dalla-Favera R
(2008) Germinal centres: Role in B-cell physiology and malignancy. Nat Rev
Immunol 8:22–33.
CrossRefMedlineWeb of Science
28. ↵
1. Ranuncolo SM,
2. et al.
(2007) Bcl-6 mediates the germinal center B cell phenotype and lymphomagenesis
through transcriptional repression of the DNA-damage sensor ATR. Nat Immun
8:705–714.
CrossRefWeb of Science
29. ↵
1. Lawrie CH,
2. et al.
(2008) MicroRNA expression in lymphocyte development and malignancy. Leukemia
22:1440–1446.
CrossRefMedlineWeb of Science
30. ↵
1. Bartkova J,
2. et al.
(2006) Oncogene-induced senescence is part of the tumorigenesis barrier imposed
by DNA damage checkpoints. Nature 444:633–637.
CrossRefMedline
31. ↵
1. Bartkova J,
2. et al.
(2005) DNA damage response as a candidate anti-cancer barrier in early human
tumorigenesis. Nature 434:864–870.
CrossRefMedline
32. ↵
1. Di Micco R,
2. et al.
(2006) Oncogene-induced senescence is a DNA damage response triggered by DNA
hyper-replication. Nature 444:638–642.
CrossRefMedline
33. ↵
1. Gorgoulis VG,
2. et al.
(2005) Activation of the DNA damage checkpoint and genomic instability in human
precancerous lesions. Nature 434:907–913.
CrossRefMedline
34. ↵
1. Herzog KH,
2. Chong MJ,
3. Kapsetaki M,
4. Morgan JI,
5. McKinnon PJ
(1998) Requirement for Atm in ionizing radiation-induced cell death in the
developing central nervous system. Science 280:1089–1091.
Abstract/FREE Full Text
35. ↵
1. Tian B,
2. Yang Q,
3. Mao Z
(2009) Phosphorylation of ATM by Cdk5 mediates DNA damage signalling and
regulates neuronal death. Nat Cell Biol 11:211–218.
CrossRefMedlineWeb of Science
36. ↵
1. Herrup K,
2. Yang Y
(2007) Cell cycle regulation in the postmitotic neuron: Oxymoron or new biology?
Nat Rev Neurosci 8:368–378.
MedlineWeb of Science
37. ↵
1. Sethupathy P,
2. Collins FS
(2008) MicroRNA target site polymorphisms and human disease. Trends Genet
24:489–497.
CrossRefMedlineWeb of Science
38. ↵
1. Mitui M,
2. et al.
(2009) Functional and computational assessment of missense variants in the ataxia-
telangiectasia mutated (ATM) gene: Mutations with increased cancer risk. Hum
Mutat 30:12–21.
CrossRefMedlineWeb of Science
39. ↵
1. Du L,
2. et al.
(2009) Nonaminoglycoside compounds induce readthrough of nonsense mutations. J
Exp Med 206:2285–2297.
Abstract/FREE Full Text
40. ↵
1. Beierle EA,
2. et al.
(2007) N-MYC regulates focal adhesion kinase expression in human neuroblastoma. J
Biol Chem 282:12503–12516.
Abstract/FREE Full Text
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