Conformation of CCAAT/Enhancer-binding Protein α Dimers Varies with Intranuclear Location in Living Cells*
Conformation of CCAAT/Enhancer-binding Protein α Dimers Varies with Intranuclear Location in Living Cells*
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CCAAT/增强子结合蛋白 α 二聚体的构象随活细胞核内位置的不同而变化*
DOI：
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发表时间：
2003
影响因子：
4.8
通讯作者：
R. Day
中科院分区：
文献类型：
--
作者：
F. Schaufele;Xia Wang;Xiaowei Liu;R. Day
蛋白质的结构决定了其生化特性，但细胞内位置和环境对蛋白质结构的影响仍不清楚。 CCAAT/增强子结合蛋白 α (C/EBPα) 是转录和细胞增殖的主要调节因子，在小鼠细胞核中转录静止的着丝粒周围区域集中并保持失活。根据连接到不同 C/EBPα 结构域的绿色荧光蛋白衍生物之间转移的荧光能量，测量活细胞中的 C/EBPα 二聚体结构。比较细胞核的着丝粒周围和非着丝粒周围区域的荧光共振能量转移水平表明，与细胞核的常染色质区域相比，C/EBPα二聚体的DNA结合结构域在着丝粒周围异染色质处距离更远并且相互作用更差。相反，转录激活域的位置和相互作用在整个细胞核中是相似的。佛波酯处理导致转录激活结构域相对于 DNA 结合结构域的位置发生变化。因此，C/EBPα 构象随核内位置和细胞环境而变化。这些“荧光共振能量转移纳米镜”技术将广泛适用于将构象和动力学变化与动态细胞内环境中 C/EBPα 或任何蛋白质的亚区室特异性作用相关联。
The structure of a protein defines its biochemical properties, but the impact of intracellular location and environment on protein structure remains poorly defined. CCAAT/enhancer-binding protein α (C/EBPα) is a master regulator of transcription and cellular proliferation that concentrates and is kept inactive at transcriptionally quiescent, pericentromeric regions in mouse cell nuclei. C/EBPα dimer structure was measured in living cells from the amounts of fluorescence energy transferred between derivatives of the green fluorescent protein attached to different C/EBPα domains. Comparing the levels of fluorescence resonance energy transfer at pericentromeric and nonpericentromeric regions of the nucleus indicated that the DNA binding domains of C/EBPα dimers were further apart and interacted more poorly at pericentromeric heterochromatin than in the more euchromatic regions of the nucleus. In contrast, the position and interactions of the transcriptional activation domains were similar throughout the nucleus. Phorbol ester treatment caused a shift in the position of the transcriptional activation domain relative to the DNA binding domain. Thus, C/EBPα conformation varies with intranuclear location and with cellular environment. These “fluorescence resonance energy transfer nanoscopy” techniques will be broadly applicable for associating conformational and kinetic variations to subcompartment-specific actions of C/EBPα or any protein in the dynamic intracellular environment.
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DOI：
10.1073/pnas.95.2.657
发表时间：
1998-01-20
影响因子：
11.1
作者：
Klug, CA;Morrison, SJ;Weissman, IL
通讯作者：
Weissman, IL
影响因子：
10.5
作者：
LEW, D;BRADY, H;MELLON, PL
通讯作者：
MELLON, PL
影响因子：
3.4
作者：
Gordon, GW;Berry, G;Herman, B
通讯作者：
Herman, B
影响因子：
10.5
作者：
Cobb, BS;Morales-Alcelay, S;Smale, ST
通讯作者：
Smale, ST
生长激素基因表达的遗传学。
影响因子：
--
作者：
Parks,JS;Abdul-Latif,H;Kinoshita,E;Meacham,LR;Pfaffle,RW;Brown,MR
通讯作者：
Brown,MR