Upstream Open Reading Frames Located in the Leader of Protein Kinase Mζ mRNA Regulate Its Translation.
Upstream Open Reading Frames Located in the Leader of Protein Kinase Mζ mRNA Regulate Its Translation.
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DOI：
10.3389/fnmol.2016.00103
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发表时间：
2016
影响因子：
4.8
通讯作者：
Kolosov P
中科院分区：
文献类型：
--
作者：
Bal NV;Susorov D;Chesnokova E;Kasianov A;Mikhailova T;Alkalaeva E;Balaban PM;Kolosov P
对于局部发生在树突中的蛋白质合成，翻译控制机制对于神经元功能比转录水平重要得多。在此，我们发现5′非翻译区（5′UTR）的uORF（上游开放阅读框）在调节蛋白激酶M β（PKM β）的翻译中起关键作用。在体外和大鼠海马神经元的原代培养物中，这些uORF的消除激活了报告蛋白的翻译。使用无细胞翻译系统，我们证明，翻译起始复合物只形成uORF。此外，我们解决的翻译抑制PKM β翻译的机制，通过uORF。我们观察到在印防己毒素的非特异性激活过程中，在神经元培养物中PKM β前导序列控制下的报告蛋白的翻译增加。我们还表明，这种机制与细胞应激中观察到的机制相似，因为将亚砷酸钠应用于神经元培养物会诱导携带PKMζ 5 'UTR的mRNA的翻译，类似于印防己毒素的激活。因此，我们假设eIF2a的磷酸化，像在细胞应激中一样，是PKM β翻译的主要调节因子。总之，我们的研究结果大大扩展了我们对uORF在活化神经元中调节PKM β翻译的作用的理解，这在LTP的早期阶段很重要。
For protein synthesis that occurs locally in dendrites, the translational control mechanisms are much more important for neuronal functioning than the transcription levels. Here, we show that uORFs (upstream open reading frames) in the 5′ untranslated region (5′UTR) play a critical role in regulation of the translation of protein kinase Mζ (PKMζ). Elimination of these uORFs activates translation of the reporter protein in vitro and in primary cultures of rat hippocampal neurons. Using cell-free translation systems, we demonstrate that translational initiation complexes are formed only on uORFs. Further, we address the mechanism of translational repression of PKMζ translation, by uORFs. We observed an increase in translation of the reporter protein under the control of PKMζ leader in neuronal culture during non-specific activation by picrotoxin. We also show that such a mechanism is similar to the mechanism seen in cell stress, as application of sodium arsenite to neuron cultures induced translation of mRNA carrying PKMζ 5′UTR similarly to picrotoxin activation. Therefore, we suppose that phosphorylation of eIF2a, like in cell stress, is a main regulator of PKMζ translation. Altogether, our findings considerably extend our understanding of the role of uORF in regulation of PKMζ translation in activated neurons, important at early stages of LTP.
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影响因子：
7.7
作者：
Andreev DE;O'Connor PB;Fahey C;Kenny EM;Terenin IM;Dmitriev SE;Cormican P;Morris DW;Shatsky IN;Baranov PV
通讯作者：
Baranov PV
影响因子：
5.3
作者：
Dmitriev, Sergey E.;Andreev, Dmitri E.;Shatsky, Ivan N.
通讯作者：
Shatsky, Ivan N.
影响因子：
5.3
作者：
Lin, Daisy;Pestova, Tatyana V.;Tiedge, Henri
通讯作者：
Tiedge, Henri
影响因子：
16
作者：
Harding, HP;Novoa, I;Ron, D
通讯作者：
Ron, D
影响因子：
5.3
作者：
Balaban, Pavel M.;Roshchin, Matvey;Malyshev, Aleksey Y.
通讯作者：
Malyshev, Aleksey Y.