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 Previous Article

Molecular and Cellular Biology, July 2003, p. 5107-5112, Vol. 23, No. 14
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.14.5107-5112.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Requirement of Watson-Crick Hydrogen Bonding for DNA Synthesis by Yeast DNA Polymerase {eta}

M. Todd Washington,1 Sandra A. Helquist,2 Eric T. Kool,2 Louise Prakash,1 and Satya Prakash1*

Sealy Center for Molecular Science, University of Texas Medical Branch at Galveston, Galveston, Texas 77555-1061,1 Department of Chemistry, Stanford University, Stanford, California 94305-50802

Received 19 March 2003/ Accepted 25 April 2003

Classical high-fidelity DNA polymerases discriminate between the correct and incorrect nucleotides by using geometric constraints imposed by the tight fit of the active site with the incipient base pair. Consequently, Watson-Crick (W-C) hydrogen bonding between the bases is not required for the efficiency and accuracy of DNA synthesis by these polymerases. DNA polymerase {eta} (Pol{eta}) is a low-fidelity enzyme able to replicate through DNA lesions. Using difluorotoluene, a nonpolar isosteric analog of thymine unable to form W-C hydrogen bonds with adenine, we found that the efficiency and accuracy of nucleotide incorporation by Pol{eta} are severely impaired. From these observations, we suggest that W-C hydrogen bonding is required for DNA synthesis by Pol{eta}; in this regard, Pol{eta} differs strikingly from classical high-fidelity DNA polymerases.

* Corresponding author. Mailing address: Sealy Center for Molecular Science, University of Texas Medical Branch at Galveston, 6.104 Blocker Medical Research Building, 11th and Mechanic Streets, Galveston, TX 77555-1061. Phone: (409) 747-8602. Fax: (409) 747-8608. E-mail: s.prakash{at}utmb.edu.

Molecular and Cellular Biology, July 2003, p. 5107-5112, Vol. 23, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.14.5107-5112.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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