What is the clinical evidence from this study?

Study design: Other. Population: Coxsackievirus B3 infection (basic science). Intervention: Mutagenesis of residue 5 in Coxsackievirus B3 3D pol vs. Wild-type 3D pol. Primary outcome: Polymerase elongation activity.

What question did this study set out to answer?

Determine the crystal structure of coxsackievirus B3 3D polymerase and characterize the functional role of the conserved residue 5 conformation across picornaviruses.

July 17, 2008Journal of Virology

Crystal Structure of Coxsackievirus B3 3D pol Highlights the Functional Importance of Residue 5 in Picornavirus Polymerases

View Full Paper

Ask AI

Bookmark

Key result

Mutation of residue 5 in coxsackievirus B3 polymerase demonstrated that elongation activity correlates with residue hydrophobicity, with the F5W mutant increasing activity to 160% of wild-type.

Population

Coxsackievirus B3 polymerase

Design

Preclinical

Authors

GCGrace CampagnolaColorado State University MWMark WeygandtColorado State University KSKirsten E. ScogginUniversity of Kentucky

Discussion

Member takes

Overview

Residue 5 hydrophobicity modulates CVB3 polymerase in vitro; leaves open in vivo effects on replication or myocarditis pathogenesis.

Key Points

Determine the crystal structure of coxsackievirus B3 3D polymerase and characterize the functional role of the conserved residue 5 conformation across picornaviruses.
Solved the X-ray crystal structure of coxsackievirus B3 3D polymerase at 2.25-Å resolution.
Performed comparative structural analyses against polymerases from poliovirus, rhinovirus, and foot-and-mouth disease virus.
Assayed coxsackievirus B3 polymerase elongation activity across residue 5 variants with varying hydrophobicity.
The 2.25-Å crystal structure revealed high homology to other picornaviral polymerases and identified a conserved distortion centered at residue 5 within the N-terminal beta-strand.
Polymerase elongation activity correlated directly with the hydrophobicity of residue 5, where substitutions with higher hydrophobicity produced greater enzymatic activity.
Structural modeling showed that residue 5 becomes buried during pre-phosphoryl transfer nucleotide repositioning, indicating the buried N-terminus stabilizes this conformational transition.

Structured PICO

Population

Coxsackievirus B3 polymerase

Intervention

Structural analysis and mutation of residue 5

Outcome

Crystal structure at 2.25-A resolution and elongation activitysurrogate

The crystal structure of coxsackievirus B3 polymerase reveals that the hydrophobicity of residue 5 is critical for elongation activity, likely by stabilizing the structure during conformational changes.

Limitations

The structure of an active picornaviral polymerase elongation complex that explicitly shows the conformational changes associated with nucleotide repositioning for catalysis remains elusive.

Cite This Study

Campagnola et al. (2008) studied Coxsackievirus B3 infection (basic science). Mutagenesis of residue 5 in Coxsackievirus B3 3D pol vs. Wild-type 3D pol was evaluated on Polymerase elongation activity. Mutation of residue 5 in coxsackievirus B3 polymerase demonstrated that elongation activity correlates with residue hydrophobicity, with the F5W mutant increasing activity to 160% of wild-type.

synapsesocial.com/papers/6a9a30ef2b9869bfd24e1eec https://doi.org/10.1128/jvi.00647-08

View Full Paper

Ask AI

Bookmark

July 17, 2008Journal of Virology

Crystal Structure of Coxsackievirus B3 3D pol Highlights the Functional Importance of Residue 5 in Picornavirus Polymerases

View Full Paper

Ask AI

Bookmark

Key result

Mutation of residue 5 in coxsackievirus B3 polymerase demonstrated that elongation activity correlates with residue hydrophobicity, with the F5W mutant increasing activity to 160% of wild-type.

Population

Coxsackievirus B3 polymerase

Design

Preclinical

Authors

GCGrace CampagnolaColorado State University MWMark WeygandtColorado State University KSKirsten E. ScogginUniversity of Kentucky

Discussion

Member takes

Overview

Residue 5 hydrophobicity modulates CVB3 polymerase in vitro; leaves open in vivo effects on replication or myocarditis pathogenesis.

Key Points

Determine the crystal structure of coxsackievirus B3 3D polymerase and characterize the functional role of the conserved residue 5 conformation across picornaviruses.
Solved the X-ray crystal structure of coxsackievirus B3 3D polymerase at 2.25-Å resolution.
Performed comparative structural analyses against polymerases from poliovirus, rhinovirus, and foot-and-mouth disease virus.
Assayed coxsackievirus B3 polymerase elongation activity across residue 5 variants with varying hydrophobicity.
The 2.25-Å crystal structure revealed high homology to other picornaviral polymerases and identified a conserved distortion centered at residue 5 within the N-terminal beta-strand.
Polymerase elongation activity correlated directly with the hydrophobicity of residue 5, where substitutions with higher hydrophobicity produced greater enzymatic activity.
Structural modeling showed that residue 5 becomes buried during pre-phosphoryl transfer nucleotide repositioning, indicating the buried N-terminus stabilizes this conformational transition.

Structured PICO

Population

Coxsackievirus B3 polymerase

Intervention

Structural analysis and mutation of residue 5

Outcome

Crystal structure at 2.25-A resolution and elongation activitysurrogate

Limitations

The structure of an active picornaviral polymerase elongation complex that explicitly shows the conformational changes associated with nucleotide repositioning for catalysis remains elusive.

Cite This Study

synapsesocial.com/papers/6a9a30ef2b9869bfd24e1eec https://doi.org/10.1128/jvi.00647-08

Also Consider

Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context:

View Full Paper

Ask AI

Bookmark