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Functional circularity of legitimate Qbeta replicase templates.

Victor I Ugarov¹, Alexander B Chetverin

¹Institute of Protein Research of the Russian Academy of Sciences, Pushchino, Moscow Region, 142290 Russia.

Journal of Molecular Biology

|May 10, 2008

Summary

Qbeta replicase efficiently amplifies RNA templates through a cooperative interaction between template termini. Disrupting this cooperation impairs RNA amplification, highlighting the functional circularity of legitimate templates.

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Area of Science:

Molecular Biology
Virology
Biochemistry

Background:

Qbeta replicase (RNA-directed RNA polymerase) amplifies specific RNAs in vitro.
Only a small fraction of RNAs serve as 'legitimate' templates for Qbeta replicase.
Template recognition involves guanosine 5'-triphosphate (GTP)-dependent initiation for legitimate templates.

Purpose of the Study:

To investigate the role of template termini cooperation in Qbeta replicase initiation and elongation.
To understand how template structure influences the formation of stable replicative complexes.
To explore the concept of functional circularity in RNA templates.

Main Methods:

Studied Qbeta replicase activity in vitro.
Utilized aurintricarboxylic acid to differentiate between stable and unstable replicative complexes.
Introduced template fragments and point mutations at the 5' terminus to disrupt cooperation.

Main Results:

Cooperation between 3' and 5' termini is crucial for GTP-dependent initiation and stable complex formation.
Disruption of terminal cooperation reduced initiation rate and yield, increased GTP requirement, and decreased overall copying rate.
Mutated or fragmented templates formed unstable postinitiation complexes.

Conclusions:

Functional circularity of legitimate Qbeta replicase templates is supported by terminal cooperation.
This cooperation is essential for efficient and stable RNA amplification.
Functional circularity may be a common feature of RNA templates in biological systems.