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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

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Published on: March 25, 2020

Relaxed Substrate Specificity in Qβ Replicase through Long-Term In Vitro Evolution.

Kohtoh Yukawa¹, Ryo Mizuuchi^2,3, Norikazu Ichihashi^1,3,4

¹Department of Life Science, Graduate School of Arts and Science, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.

Life (Basel, Switzerland)

|January 21, 2022

Summary

Early life

Keywords:

Qβ replicase RNA world experimental evolution origin of DNA

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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

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Published on: March 16, 2011

Area of Science:

Evolutionary biology
Molecular evolution
Origin of life

Background:

The transition from RNA to DNA is a key event in early life evolution.
Understanding how replication enzymes adapted is crucial for evolutionary history.

Purpose of the Study:

To investigate how RNA replication enzymes might have adapted to incorporate deoxyribonucleotide triphosphates (dNTPs).
To explore the plausibility of an easier RNA to DNA transition.

Main Methods:

Evolving an RNA replication enzyme in the absence of dNTPs.
Assessing the enzyme's substrate specificity and incorporation of labeled dNTPs.

Main Results:

The evolved RNA replication enzyme showed relaxed substrate specificity.
The enzyme successfully incorporated dNTPs, indicating a potential for DNA synthesis.

Conclusions:

Ancient RNA replication enzymes could have incorporated dNTPs once they became available.
The RNA to DNA transition may have been more facile than previously assumed.