Journal of Molecular Evolution

, Volume 42, Issue 5, pp 493–499 | Cite as

De Novo synthesis of DNA-like molecules by polynucleotide phosphorylase in vitro

  • Mirko Beljanski
Articles

Abstract

In the presence of Mg2+ ions, polynucleotide phosphorylase (PNPase, EC 2.7.7.8) is known to synthesize RNA-like polymers using ribonucleoside-5′-diphosphate (NDP) substrates but to be unable to utilize deoxyribonucleoside substrates. Our experiments show that when MgCl2 is replaced by FeCl3, PNPase becomes able to synthesize deoxyheteropolymers using deoxyribonucleoside-5′-diphosphates (dNDPs). The deoxyheteropolymer formed from the four dNDPs is degraded by pancreatic DNase, but not by RNase, and is readily used as a template by DNA-dependent DNA polymerase. Synthesis of this DNA-like polymer is accomplished de novo without the help of any primer or preexisting template. What is more, dA/dG and dC/dT ratios of polymers synthesized by different bacterial PNPases closely match ratios found in DNA of the bacterial species the enzyme came from.

Key words

Polynucleotide phosphorylase De novo deoxypolymer synthesis DNA Deoxyribonucleoside-5′-diphosphates Ferric ions 

Abbreviations

PNPase

polynucleotide phosphorylase

dNDP

deoxyribonucleoside-5′-diphosphates

NDP

ribonucleoside-5′-diphosphates

DNA

deoxyribonucleic acid

RNA

ribonucleic acid

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Copyright information

© Springer-Verlag New York Inc 1996

Authors and Affiliations

  • Mirko Beljanski
    • 1
  1. 1.Cerbiol ApplicationCentre de Recherche BiologiqueSaint-PrimFrance

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