Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Applied Bio

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Science 30 September 2005:
Vol. 309. no. 5744, pp. 2219 - 2222
DOI: 10.1126/science.1116336
Prev | Table of Contents | Next

Reports

Rev1 Employs a Novel Mechanism of DNA Synthesis Using a Protein Template

Deepak T. Nair,1 Robert E. Johnson,2 Louise Prakash,2 Satya Prakash,2 Aneel K. Aggarwal1*

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.

1 Structural Biology Program, Department of Physiology and Biophysics, Mount Sinai School of Medicine, Box 1677, 1425 Madison Avenue, New York, NY 10029, USA.
2 Sealy Center for Molecular Science, University of Texas Medical Branch, 6.014 Medical Research Building, 11th and Mechanic Streets, Galveston, TX 77755–1061, USA.

* To whom correspondence should be addressed. E-mail: aggarwal{at}inka.mssm.edu

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Kinetic Analysis of Translesion Synthesis Opposite Bulky N2- and O6-Alkylguanine DNA Adducts by Human DNA Polymerase REV1.
J.-Y. Choi and F. P. Guengerich (2008)
J. Biol. Chem. 283, 23645-23655
   Abstract »    Full Text »    PDF »
Role for DNA Polymerase {kappa} in the Processing of N2-N2-Guanine Interstrand Cross-links.
I. G. Minko, M. B. Harbut, I. D. Kozekov, A. Kozekova, P. M. Jakobs, S. B. Olson, R. E. Moses, T. M. Harris, C. J. Rizzo, and R. S. Lloyd (2008)
J. Biol. Chem. 283, 17075-17082
   Abstract »    Full Text »    PDF »
Chemical mapping of cytosines enzymatically flipped out of the DNA helix.
D. Daujotyte, Z. Liutkeviciute, G. Tamulaitis, and S. Klimasauskas (2008)
Nucleic Acids Res. 36, e57
   Abstract »    Full Text »    PDF »
Mechanistic consequences of temperature on DNA polymerization catalyzed by a Y-family DNA polymerase.
K. A. Fiala, S. M. Sherrer, J. A. Brown, and Z. Suo (2008)
Nucleic Acids Res. 36, 1990-2001
   Abstract »    Full Text »    PDF »
Complex Formation of Yeast Rev1 with DNA Polymerase {eta}.
N. Acharya, L. Haracska, S. Prakash, and L. Prakash (2007)
Mol. Cell. Biol. 27, 8401-8408
   Abstract »    Full Text »    PDF »
AtREV1, a Y-Family DNA Polymerase in Arabidopsis, Has Deoxynucleotidyl Transferase Activity in Vitro.
S. Takahashi, A. N. Sakamoto, A. Tanaka, and K. Shimizu (2007)
Plant Physiology 145, 1052-1060
   Abstract »    Full Text »    PDF »
What a difference a decade makes: Insights into translesion DNA synthesis.
W. Yang and R. Woodgate (2007)
PNAS 104, 15591-15598
   Abstract »    Full Text »    PDF »
A Ubiquitin-binding Motif in the Translesion DNA Polymerase Rev1 Mediates Its Essential Functional Interaction with Ubiquitinated Proliferating Cell Nuclear Antigen in Response to DNA Damage.
A. Wood, P. Garg, and P. M. J. Burgers (2007)
J. Biol. Chem. 282, 20256-20263
   Abstract »    Full Text »    PDF »
Sloppy Bypass of an Abasic Lesion Catalyzed by a Y-family DNA Polymerase.
K. A. Fiala and Z. Suo (2007)
J. Biol. Chem. 282, 8199-8206
   Abstract »    Full Text »    PDF »
Complex Formation with Rev1 Enhances the Proficiency of Saccharomyces cerevisiae DNA Polymerase {zeta} for Mismatch Extension and for Extension Opposite from DNA Lesions.
N. Acharya, R. E. Johnson, S. Prakash, and L. Prakash (2006)
Mol. Cell. Biol. 26, 9555-9563
   Abstract »    Full Text »    PDF »
Novel Role for the C Terminus of Saccharomyces cerevisiae Rev1 in Mediating Protein-Protein Interactions.
S. D'Souza and G. C. Walker (2006)
Mol. Cell. Biol. 26, 8173-8182
   Abstract »    Full Text »    PDF »
Human DNA polymerase {kappa} forms nonproductive complexes with matched primer termini but not with mismatched primer termini.
K. D. Carlson, R. E. Johnson, L. Prakash, S. Prakash, and M. T. Washington (2006)
PNAS 103, 15776-15781
   Abstract »    Full Text »    PDF »
Role of Single-stranded DNA in Targeting REV1 to Primer Termini.
Y. Masuda and K. Kamiya (2006)
J. Biol. Chem. 281, 24314-24321
   Abstract »    Full Text »    PDF »
The critical mutagenic translesion DNA polymerase Rev1 is highly expressed during G2/M phase rather than S phase.
L. S. Waters and G. C. Walker (2006)
PNAS 103, 8971-8976
   Abstract »    Full Text »    PDF »
Kinetic Evidence for Inefficient and Error-prone Bypass across Bulky N2-Guanine DNA Adducts by Human DNA Polymerase {iota}.
J.-Y. Choi and F. P. Guengerich (2006)
J. Biol. Chem. 281, 12315-12324
   Abstract »    Full Text »    PDF »
Ubiquitylation of yeast proliferating cell nuclear antigen and its implications for translesion DNA synthesis.
L. Haracska, I. Unk, L. Prakash, and S. Prakash (2006)
PNAS 103, 6477-6482
   Abstract »    Full Text »    PDF »
Saccharomyces cerevisiae Polymerase {zeta} Functions in Mitochondria.
H. Zhang, A. Chatterjee, and K. K. Singh (2006)
Genetics 172, 2683-2688
   Abstract »    Full Text »    PDF »
DNA nicking by HinP1I endonuclease: bending, base flipping and minor groove expansion.
J. R. Horton, X. Zhang, R. Maunus, Z. Yang, G. G. Wilson, R. J. Roberts, and X. Cheng (2006)
Nucleic Acids Res. 34, 939-948
   Abstract »    Full Text »    PDF »
A new anti conformation for N-(deoxyguanosin-8-yl)-2-acetylaminofluorene (AAF-dG) allows Watson-Crick pairing in the Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4).
L. Wang and S. Broyde (2006)
Nucleic Acids Res. 34, 785-795
   Abstract »    Full Text »    PDF »
Polymerase with a protein template.
N. LeBrasseur (2005)
J. Cell Biol. 171, 192
   Full Text »    PDF »


ADVERTISEMENT
Click Me!

ADVERTISEMENT
Click Me!

To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)