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Updated: Jan 8, 2026

In Vitro Transcribed RNA-based Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-infected Cells

In Vitro Transcribed RNA-based Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-infected Cells

Published on: May 1, 2019

HCVコーディング領域におけるリボソーム結合型シュードノットがウイルス翻訳を調整することによりウイルス増殖を促進する

Han Wan¹, Harim Jang², Ling Xu²

¹Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06511, USA.

|December 21, 2025

Abstract:

Recent studies have uncovered a number of functional RNA structures in RNA viruses, yet their regulatory roles remain poorly understood. Here, using an unbiased proteomic approach alongside targeted biochemical assays, we investigate a previously uncharacterized functional pseudoknot (pk1) within the hepatitis C virus coding region and show that it stably interacts with host ribosomes, inhibiting translation and potentially acting as a regulator between viral translation and RNA genome replication. Comparative structural analysis identifies pk1-like elements in related RNA viruses, suggesting a conserved regulatory mechanism. This study expands the known functions of pseudoknots in viral coding regions beyond frameshifting, highlights the critical role of RNA structure-mediated regulation within viral open reading frames, and provides insight into the design of viral therapeutics and vaccines.

キーワード:

微生物学分子生物学 RNA構造 RNAウイルス RNA結合タンパク質アンチセンスRNA 遺伝暗号ロックド核酸リボレギュレーション翻訳阻害翻訳開始

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A Protocol for Analyzing Hepatitis C Virus Replication

A Protocol for Analyzing Hepatitis C Virus Replication

Published on: June 26, 2014

Two Methods of Heterokaryon Formation to Discover HCV Restriction Factors

Two Methods of Heterokaryon Formation to Discover HCV Restriction Factors

Published on: July 16, 2012

関連する概念動画

Leaky Scanning

Leaky Scanning

During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R...

Initiation of Translation

Initiation of Translation

Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...

Initiation of Translation

Initiation of Translation

Ribosomal RNA Synthesis

Ribosomal RNA Synthesis

Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...

Ribosomal RNA Synthesis

Ribosomal RNA Synthesis

Improving Translational Accuracy

Improving Translational Accuracy

Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...