Site-selective incorporation of phosphorylated tyrosine into the p50 subunit of NF-κB and activation of its downstream gene CD40.
Site-selective incorporation of phosphorylated tyrosine into the p50 subunit of NF-κB and activation of its downstream gene CD40.
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DOI：
10.1039/d1cc04726d
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发表时间：
2021-11-25
期刊：
影响因子：
--
通讯作者：
Hecht SM
中科院分区：
文献类型：
--
作者：
Chen S;Ji X;Dedkova LM;Hecht SM
核因子-κB转录激活剂家族负责大量基因的表达，这些基因控制着细胞发育和生存等关键功能。P50亚基已经被广泛研究，已知包括13种酪氨酸，但文献中还没有定义伴随p50功能的酪氨酸磷酸化的程度和模式，特别是在基因表达的选择性水平上。在这项研究中，利用含有修饰核糖体的大肠杆菌体外表达系统，将磷酸化酪氨酸(PTyr)选择性地掺入p50亚基中。在人类T细胞中，在p50的60或82位含有pTyr的核因子-κB强烈地增加了CD40的表达，这是癌症或病毒免疫治疗的潜在靶点。研究了CD40启动子与启动子的结合，证实了NF-κB p50/p65异源二聚体中的两个pTyr残基促进了这一过程，并支持磷酸化化学计量学的可能重要性。本研究为研究酪氨酸残基的磷酸化改变蛋白质与DNA启动子的结合提供了一种新的方法，并有助于DNA表达的方便性。核因子-DNABp50亚基关键酪氨酸的磷酸化在体外增强了κ启动子的结合，随后在体内增强了蛋白质的翻译。
The NF-κB family of transcriptional activators is responsible for the expression of numerous genes that control key functions such as cell development and survival. Subunit p50 has been studied extensively and is known to include 13 tyrosines, but the extent and pattern of tyrosine phosphorylation that accompanies p50 function has not been defined in the literature, especially at the level of selectivity of gene expression. In this study, phosphorylated tyrosine (pTyr) was site-selectively incorporated into the p50 subunit using an E. coli in vitro expression system containing a modified ribosome. In human T cells, the NF-κBs containing a pTyr at position 60 or 82 of p50 strongly increased the expression of CD40, which is a potential target for cancer or viral immunotherapy. Promoter DNA binding was studied for CD40 promoters, and verified two pTyr residues in NF-κB p50/p65 heterodimers that facilitated this process, and that support the possible importance of phosphorylation stoichiometry. This study defines a new approach for studying tyrosine residues whose phosphorylation alters protein binding to DNA promoters, and contributes to the facility of DNA expression. Phosphorylation of key tyrosines in the NF-κB p50 subunit enhances DNA promoter binding in vitro and subsequent protein translation in vivo.
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影响因子：
6
作者：
Christian F;Smith EL;Carmody RJ
通讯作者：
Carmody RJ
影响因子：
20.3
作者：
Hinz, M;Löser, P;Scheidereit, C
通讯作者：
Scheidereit, C
DOI：
10.1084/jem.185.12.2053
发表时间：
1997-06-16
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Carbone E;Ruggiero G;Terrazzano G;Palomba C;Manzo C;Fontana S;Spits H;Kärre K;Zappacosta S
通讯作者：
Zappacosta S
影响因子：
64.8
作者：
Chen, FE;Huang, DB;Ghosh, G
通讯作者：
Ghosh, G
影响因子：
3.8
作者：
Kang, JL;Jung, HJ;Kim, HR
通讯作者：
Kim, HR