The ATM kinase signaling induced by the low-energy β-particles emitted by (33)P is essential for the suppression of chromosome aberrations and is greater than that induced by the energetic β-particles emitted by (32)P.
The ATM kinase signaling induced by the low-energy β-particles emitted by (33)P is essential for the suppression of chromosome aberrations and is greater than that induced by the energetic β-particles emitted by (32)P.
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DOI：
10.1016/j.mrfmmm.2011.01.005
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发表时间：
2011-03-15
影响因子：
2.3
通讯作者：
Bakkenist, Christopher J.
中科院分区：
文献类型：
--
作者：
White, Jason S.;Yue, Ning;Hu, Jing;Bakkenist, Christopher J.
共济失调-毛细血管扩张突变（ATM）编码一种核丝氨酸/苏氨酸蛋白激酶，其活性在暴露于低剂量电离辐射（IR）的细胞中增加。在这里，我们研究ATM激酶激活细胞暴露于32 P-或33 P-正磷酸盐的条件下，通常采用的代谢标记实验。我们计算了在含有1 mCi高能辐射的2 ml培养基中孵育的5 cm × 5 cm单层细胞的IR吸收剂量，（1.70 MeV）β-粒子发射体32 P-正磷酸盐照射30 min的吸收剂量为~1戈伊IR。（0.24 MeV）β粒子发射体33 P-正磷酸盐的IR约为0.18戈伊。我们发现，由33 P发射的低能β粒子比由32 P发射的高能β粒子诱导更多的电离辐射诱导灶（IRIF）和更大的ATM激酶信号传导。因此，我们证明，这是不合适的，使用33 P-正磷酸盐作为32 P-正磷酸盐在实验研究DNA双链断裂（DSB）的DNA损伤反应的阴性对照。值得注意的是，我们表明，ATM积累在染色质组分时，ATM激酶活性受到抑制，在暴露于任何放射性核素。最后，我们还表明，当ATM激酶活性在暴露于~0.36戈伊的β粒子33 P发射的过程中受到抑制时，染色体畸变在细胞中积累。因此，我们建议，直接细胞暴露于33 P-正磷酸是一个很好的手段，诱导和标记IR诱导的ATM激酶依赖性磷酸化蛋白质组。
Ataxia-telangiectasia mutated (ATM) encodes a nuclear serine/threonine protein kinase whose activity is increased in cells exposed to low doses of ionizing radiation (IR). Here we examine ATM kinase activation in cells exposed to either 32P- or 33P-orthophosphate under conditions typically employed in metabolic labelling experiments. We calculate that the absorbed dose of IR delivered to a 5 cm × 5 cm monolayer of cells incubated in 2 ml media containing 1 mCi of the high-energy (1.70 MeV) β-particle emitter 32P-orthophosphate for 30 min is ~1 Gy IR. The absorbed dose of IR following an otherwise identical exposure to the low-energy (0.24 MeV) β-particle emitter 33P-orthophosphate is ~0.18 Gy IR. We show that low-energy β-particles emitted by 33P induce a greater number of ionizing radiation-induced foci (IRIF) and greater ATM kinase signaling than energetic β-particles emitted by 32P. Hence, we demonstrate that it is inappropriate to use 33P-orthophosphate as a negative control for 32P-orthophosphate in experiments investigating DNA damage responses to DNA double-strand breaks (DSBs). Significantly, we show that ATM accumulates in the chromatin fraction when ATM kinase activity is inhibited during exposure to either radionuclide. Finally, we also show that chromosome aberrations accumulate in cells when ATM kinase activity is inhibited during exposure to ~0.36 Gy β-particles emitted by 33P. We therefore propose that direct cellular exposure to 33P-orthophosphate is an excellent means to induce and label the IR-induced, ATM kinase-dependent phosphoproteome.
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影响因子：
10.5
作者：
Siliciano, JD;Canman, CE;Kastan, MB
通讯作者：
Kastan, MB
DOI：
10.1073/pnas.190030497
发表时间：
2000-09-12
影响因子：
11.1
作者：
Matsuoka, S;Rotman, G;Elledge, SJ
通讯作者：
Elledge, SJ
影响因子：
56.9
作者：
SAVITSKY, K;BARSHIRA, A;SHILOH, Y
通讯作者：
SHILOH, Y
影响因子：
7.3
作者：
Olsen, Jesper V.;Vermeulen, Michiel;Mann, Matthias
通讯作者：
Mann, Matthias
DOI：
10.1073/pnas.1002175107
发表时间：
2010-04-13
影响因子：
11.1
作者：
de Feraudy, Sebastien;Revet, Ingrid;Cleaver, James E.
通讯作者：
Cleaver, James E.