Related Experiment Video

Updated: Apr 27, 2026

Multi-target Parallel Processing Approach for Gene-to-structure Determination of the Influenza Polymerase PB2 Subunit
22:10

Multi-target Parallel Processing Approach for Gene-to-structure Determination of the Influenza Polymerase PB2 Subunit

Published on: June 28, 2013

13.8K

Secondary structure of a conserved domain in an intron of influenza A M1 mRNA

Tian Jiang1, Scott D Kennedy, Walter N Moss

  • 1Department of Chemistry and Center for RNA Biology, University of Rochester , Rochester, New York 14627, United States.

Biochemistry
|July 16, 2014
PubMed

Insights

Influenza A virus RNA structures are crucial for infection. This study confirms a predicted conserved three-way multibranch RNA structure in M1/M2 pre-mRNA, essential for virus function and potential therapeutic targets.

Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Influenza A virus relies heavily on RNA during infection, yet the functional roles of its RNA structures remain largely unexplored.
  • Bioinformatic analyses have identified potentially stable and conserved RNA structures within the influenza A virus genome.
  • A specific conserved structure was predicted in the M1/M2 protein-coding pre-mRNA, downstream of the M2 mRNA 5' splice site.

Purpose of the Study:

  • To biochemically and structurally characterize a predicted conserved RNA structure in influenza A virus M1/M2 pre-mRNA.
  • To validate the predicted three-way multibranch RNA structure using experimental methods.
  • To investigate the stability of this RNA structure under varying ionic conditions.

Main Methods:

  • Biochemical structural mapping techniques were employed to analyze the RNA.
  • Site-directed mutagenesis was used to probe structural elements.
  • Nuclear Magnetic Resonance (NMR) spectroscopy, specifically imino proton NMR, was utilized to assess secondary structure.
  • Optical melting curve analysis was performed to determine RNA stability.

Main Results:

  • The predicted three-way multibranch RNA structure was confirmed experimentally.
  • NMR analysis showed no significant change in secondary structure upon addition of magnesium ions (MgCl2) to potassium chloride (KCl).
  • Optical melting experiments indicated increased RNA stability in the presence of both NaCl and a combination of KCl and MgCl2, with melting temperatures rising approximately 14 °C compared to KCl alone.

Conclusions:

  • Experimental evidence validates the predicted conserved three-way multibranch RNA structure in influenza A virus M1/M2 pre-mRNA.
  • The RNA structure exhibits stability in the presence of physiological salt concentrations and divalent cations.
  • These findings establish a foundation for future research into the functional roles of this RNA structure and for the development of targeted therapeutics.

More Related Videos

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein09:39

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Published on: April 21, 2015

17.4K
An Integrated Approach for Microprotein Identification and Sequence Analysis09:37

An Integrated Approach for Microprotein Identification and Sequence Analysis

Published on: July 12, 2022

3.1K

Related Concept Videos

Leaky Scanning02:28

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...
4.5K
Conserved Binding Sites01:49

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
4.1K
Conservation of Protein Domains Over Different Proteins02:26

Conservation of Protein Domains Over Different Proteins

Protein domains are small structurally independent units that are part of a single amino acid chain.  Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to...
11.7K
Nucleic Acid Structure01:25

Nucleic Acid Structure

The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms  a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA...
8.0K
RNA Structure01:23

RNA Structure

Overview
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
68.8K
RNA Structure01:19

RNA Structure

The basic structure of RNA consists of a string of ribonucleotides attached by phosphodiester bonds. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA) involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three...
6.5K