相关实验视频

Updated: Jul 10, 2025

NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells

NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells

Published on: January 12, 2020

SARS-CoV-2 ORF3a介导的NF-κB激活不依赖于TRAF结合序列

Brianna M Busscher¹, Henock B Befekadu², Zhonghua Liu^1,3

¹Department of Pathology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106, USA.

|November 25, 2023

概括

SARS-CoV-2 ORF3a 蛋白可能通过与 TRAF 蛋白结合以外的机制激活 NF-κB,这是一个关键的炎症途径. 这一发现为COVID-19病原体提供了新的见解.

科学领域:

病毒学病毒学
免疫学免疫学免疫学
分子生物学分子生物学

背景情况:

严重急性呼吸系统综合征冠状病毒2 (SARS-CoV-2) 导致COVID-19,其特点是过度炎症.
SARS-CoV-2辅助蛋白ORF3a与COVID-19的病理和炎症有关.
核因子kappa B (NF-κB) 是一个关键的炎症途径.

研究的目的:

研究SARS-CoV-2ORF3a蛋白在与TNF受体关联因子 (TRAF) 蛋白结合中的作用.
确定ORF3a诱导NF-κB激活的能力.
阐明ORF3a影响炎症通路的机制.

主要方法:

用X射线结晶学分析蛋白质结构.
光极化试验,以评估蛋白质与配体的结合.
双露西法酶测定测量NF-κB激活的方法.
位点定向突变发生以研究蛋白质功能.

主要成果:

在ORF3a N-终端和TRAF蛋白之间观察到低亲和度结合.
ORF3a 显示了激活 NF-κB 的能力.
在ORF3a中对N端TRAF结合序列 (PIQAS) 发生突变并没有显著降低NF-κB激活.

结论:

在SARS-CoV-2中,ORF3a蛋白可以激活NF-κB.
通过N端PIQAS序列与TRAF蛋白的相互作用不是ORF3a诱导NF-κB激活的主要机制.
其他机制可能会调解ORF3a在激活NF-κB和促进COVID-19病理方面的作用.

关键词:

在 NF-κBB 中.在SARS-CoV-2的ORF3a.它们是TRAF蛋白质.这是一种炎症炎症炎症炎症.

更多相关视频

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3

Published on: January 24, 2016

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α

Published on: November 2, 2018

相关概念视频

NF-κB-dependent Signaling Pathway

NF-κB-dependent Signaling Pathway

The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The...

MAPK Signaling Cascades

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...

Regulation of Nuclear Protein Sorting

Regulation of Nuclear Protein Sorting

Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...

Co-activators and Co-repressors

Co-activators and Co-repressors

Gene transcription is regulated by the synergistic action of several proteins that form a complex at a gene regulatory site. This is observed in eukaryotes, where the regulation of gene expression is a complex process. Regulatory proteins in eukaryotes can broadly be classified into two types – regulators that bind directly to specific DNA sequences and co-regulators that associate with regulatory proteins but cannot directly bind to the DNA. These co-regulators are further divided into...

TGF - β Signaling Pathway

TGF - β Signaling Pathway

The TGF-β signaling pathway regulates cell growth, differentiation, adhesion, motility, and development. TGF-β ligands that induce TGF-β signaling are synthesized in their latent form. Several proteases or cell surface receptors such as integrins act upon the latent form, releasing the active ligand. There are three types of mammalian TGF-βs: (TGF-β1, TGF-β2, and TGF-β3) that bind as homodimers or heterodimers to TGF-β receptors. The TGF-β receptors...

The JAK-STAT Signaling Pathway

The JAK-STAT Signaling Pathway

Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as SH2...