Rev1 employs a novel mechanism of DNA synthesis using a protein template

Deepak T Nair; Robert E Johnson; Louise Prakash; Satya Prakash; Aneel K Aggarwal

doi:10.1126/science.1116336

Rev1 employs a novel mechanism of DNA synthesis using a protein template

Science. 2005 Sep 30;309(5744):2219-22. doi: 10.1126/science.1116336.

Authors

Deepak T Nair¹, Robert E Johnson, Louise Prakash, Satya Prakash, Aneel K Aggarwal

Affiliation

¹ Structural Biology Program, Department of Physiology and Biophysics, Mount Sinai School of Medicine, Box 1677, 1425 Madison Avenue, New York, NY 10029, USA.

PMID: 16195463
DOI: 10.1126/science.1116336

Abstract

The Rev1 DNA polymerase is highly specialized for the incorporation of C opposite template G. We present here the crystal structure of yeast Rev1 bound to template G and incoming 2'-deoxycytidine 5'-triphosphate (dCTP), which reveals that the polymerase itself dictates the identity of the incoming nucleotide, as well as the identity of the templating base. Template G and incoming dCTP do not pair with each other. Instead, the template G is evicted from the DNA helix, and it makes optimal hydrogen bonds with a segment of Rev1. Also, unlike other DNA polymerases, incoming dCTP pairs with an arginine rather than the templating base, which ensures the incorporation of dCTP over other incoming nucleotides. This mechanism provides an elegant means for promoting proficient and error-free synthesis through N2-adducted guanines that obstruct replication.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, P.H.S.

MeSH terms

Arginine / metabolism
Base Pairing
Binding Sites
Catalytic Domain
Crystallography, X-Ray
DNA Replication*
DNA, Fungal / biosynthesis*
Deoxycytosine Nucleotides / chemistry
Deoxycytosine Nucleotides / metabolism*
Guanine / chemistry
Guanine / metabolism*
Hydrogen Bonding
Hydrophobic and Hydrophilic Interactions
Models, Molecular
Nucleotidyltransferases / chemistry*
Nucleotidyltransferases / genetics
Nucleotidyltransferases / metabolism*
Protein Conformation
Protein Structure, Secondary
Protein Structure, Tertiary
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / metabolism
Saccharomyces cerevisiae / enzymology
Saccharomyces cerevisiae / metabolism
Saccharomyces cerevisiae Proteins / chemistry*
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism*
Templates, Genetic

Substances

DNA, Fungal
Deoxycytosine Nucleotides
Recombinant Fusion Proteins
Saccharomyces cerevisiae Proteins
2'-deoxycytidine 5'-triphosphate
Guanine
Arginine
Nucleotidyltransferases
REV1 protein, S cerevisiae

Associated data

PDB/2AQ4