Nucleotide Analogues as Probes for DNA and RNA Polymerases

Robert D. Kuchta1

1 University of Colorado, Boulder, Colorado
Publication Name:  Current Protocols in Chemical Biology
Unit Number:   
DOI:  10.1002/9780470559277.ch090203
Online Posting Date:  June, 2010
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Abstract

Nucleotide analogues represent a major class of anti‐cancer and anti‐viral drugs, and provide an extremely powerful tool for dissecting the mechanisms of DNA and RNA polymerases. While the basic assays themselves are relatively straightforward, a key issue is to appropriately design the studies to answer the mechanistic question of interest. This unit addresses the major issues involved in designing these studies, and some of the potential difficulties that arise in interpreting the data. Examples are given for the type of analogues typically used, the experimental approaches with different polymerases, and issues with data interpretation. Curr. Protoc. Chem Biol. 2:111‐124. © 2010 by John Wiley & Sons, Inc.

Keywords: polymerase; nucleotide; DNA; RNA; kinetics

     
 
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Table of Contents

  • Introduction
  • Sugar Analogues
  • Base Analogues
  • Effects of DNA Sequence
  • 3′‐5′ Exonuclease Activity
  • Challenges of RNA Polymerases
  • Data Interpretation
  • Examples of Difficulty Interpreting how a Polymerase Interacts with a Nucleotide
  • Acknowledgements
  • Literature Cited
  • Figures
     
 
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Materials

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Literature Cited

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