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.

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Science 17 April 1992:
Vol. 256. no. 5055, pp. 350 - 355
DOI: 10.1126/science.256.5055.350
Prev | Table of Contents | Next

Articles

Allosteric Effects of Nucleotide Cofactors on Escherichia coli Rep Helicase&DNA Binding

Isaac Wong 1 and Timothy M. Lohman 1

1 Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110

The Escherichia coli Rep helicase unwinds duplex DNA during replication. The functional helicase appears to be a dimer that forms only on binding DNA. Both protomers of the dimer can bind either single-stranded or duplex DNA. Because binding and hydrolysis of adenosine triphosphate (ATP) are essential for helicase function, the energetics of DNA binding and DNA-induced Rep dimerization were studied quantitatively in the presence of the nucleotide cofactors adenosine diphosphate (ADP) and the nonhydrolyzable ATP analog AMPP(NH) P. Large allosteric effects of nucleotide cofactors on DNA binding to Rep were observed. Binding of ADP favored Rep dimers in which both protomers bound singlestranded DNA, whereas binding of AMPP(NH)P favored simultaneous binding of both single-stranded and duplex DNA to the Rep dimer. A rolling model for the active unwinding of duplex DNA by the dimeric Rep helicase is proposed that explains vectorial unwinding and predicts that helicase translocation along DNA is coupled to ATP binding, whereas ATP hydrolysis drives unwinding of multiple DNA base pairs for each catalytic event.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Elucidating the mechanism of DNA-dependent ATP hydrolysis mediated by DNA-dependent ATPase A, a member of the SWI2/SNF2 protein family.
M. Nongkhlaw, P. Dutta, J. W. Hockensmith, S. S. Komath, and R. Muthuswami (2009)
Nucleic Acids Res. 37, 3332-3341
   Abstract »    Full Text »    PDF »
Mechanisms of Helicases.
S. S. Patel and I. Donmez (2006)
J. Biol. Chem. 281, 18265-18268
   Full Text »    PDF »
Escherichia coli RecQ Is a Rapid, Efficient, and Monomeric Helicase.
X.-D. Zhang, S.-X. Dou, P. Xie, J.-S. Hu, P.-Y. Wang, and X. G. Xi (2006)
J. Biol. Chem. 281, 12655-12663
   Abstract »    Full Text »    PDF »
The DNA Binding Properties of the Escherichia coli RecQ Helicase.
S.-X. Dou, P.-Y. Wang, H. Q. Xu, and X. G. Xi (2004)
J. Biol. Chem. 279, 6354-6363
   Abstract »    Full Text »    PDF »
The Nonstructural Protein 3 Protease/Helicase Requires an Intact Protease Domain to Unwind Duplex RNA Efficiently.
D. N. Frick, R. S. Rypma, A. M. I. Lam, and B. Gu (2004)
J. Biol. Chem. 279, 1269-1280
   Abstract »    Full Text »    PDF »
Adenosine 5'-O-(3-thio)triphosphate (ATP{gamma}S) is a substrate for the nucleotide hydrolysis and RNA unwinding activities of eukaryotic translation initiation factor eIF4A.
M. L. PECK and D. HERSCHLAG (2003)
RNA 9, 1180-1187
   Abstract »    Full Text »    PDF »
Plant DNA helicases: the long unwinding road.
N. Tuteja (2003)
J. Exp. Bot. 54, 2201-2214
   Abstract »    Full Text »    PDF »
The Escherichia coli RecQ Helicase Functions as a Monomer.
H. Q. Xu, E. Deprez, A. H. Zhang, P. Tauc, M. M. Ladjimi, J.-C. Brochon, C. Auclair, and X. G. Xi (2003)
J. Biol. Chem. 278, 34925-34933
   Abstract »    Full Text »    PDF »
Phospholipid-induced Monomerization and Signal-peptide-induced Oligomerization of SecA.
J. Benach, Y.-T. Chou, J. J. Fak, A. Itkin, D. D. Nicolae, P. C. Smith, G. Wittrock, D. L. Floyd, C. M. Golsaz, L. M. Gierasch, et al. (2003)
J. Biol. Chem. 278, 3628-3638
   Abstract »    Full Text »    PDF »
The 2B domain of the Escherichia coli Rep protein is not required for DNA helicase activity.
W. Cheng, K. M. Brendza, G. H. Gauss, S. Korolev, G. Waksman, and T. M. Lohman (2002)
PNAS 99, 16006-16011
   Abstract »    Full Text »    PDF »
Helicase from Hepatitis C Virus, Energetics of DNA Binding.
M. K. Levin and S. S. Patel (2002)
J. Biol. Chem. 277, 29377-29385
   Abstract »    Full Text »    PDF »
Modularity and Specialization in Superfamily 1 and 2 Helicases.
M. R. Singleton and D. B. Wigley (2002)
J. Bacteriol. 184, 1819-1826
   Full Text »    PDF »
Distinguishing Inchworm and Hand-Over-Hand Processive Kinesin Movement by Neck Rotation Measurements.
W. Hua, J. Chung, and J. Gelles (2002)
Science 295, 844-848
   Abstract »    Full Text »    PDF »
DNA and ATP Binding Activities of the Baculovirus DNA Helicase P143.
V. V. McDougal and L. A. Guarino (2001)
J. Virol. 75, 7206-7209
   Abstract »    Full Text »
Defining the roles of individual residues in the single-stranded DNA binding site of PcrA helicase.
M. S. Dillingham, P. Soultanas, P. Wiley, M. R. Webb, and D. B. Wigley (2001)
PNAS 98, 8381-8387
   Abstract »    Full Text »    PDF »
Unwinding of nucleic acids by HCV NS3 helicase is sensitive to the structure of the duplex.
A. J. Tackett, L. Wei, C. E. Cameron, and K. D. Raney (2001)
Nucleic Acids Res. 29, 565-572
   Abstract »    Full Text »    PDF »
Characterisation of the catalytically active form of RecG helicase.
P. McGlynn, A. A. Mahdi, and R. G. Lloyd (2000)
Nucleic Acids Res. 28, 2324-2332
   Abstract »    Full Text »    PDF »
Enzymatic properties of hepatitis C virus NS3-associated helicase.
C. Paolini, R. De Francesco, and P. Gallinari (2000)
J. Gen. Virol. 81, 1335-1345
   Abstract »    Full Text »
Reconstitution of a minimal RNA degradosome demonstrates functional coordination between a 3' exonuclease and a DEAD-box RNA helicase.
G. A. Coburn, X. Miao, D. J. Briant, and G. A. Mackie (1999)
Genes & Dev. 13, 2594-2603
   Abstract »    Full Text »
Ded1p, a DEAD-box Protein Required for Translation Initiation in Saccharomyces cerevisiae, Is an RNA Helicase.
I. Iost, M. Dreyfus, and P. Linder (1999)
J. Biol. Chem. 274, 17677-17683
   Abstract »    Full Text »    PDF »
Escherichia coli DNA Helicase II Is Active as a Monomer.
L. E. Mechanic, M. C. Hall, and S. W. Matson (1999)
J. Biol. Chem. 274, 12488-12498
   Abstract »    Full Text »    PDF »
A Region Near the C-Terminal End of Escherichia coli DNA Helicase II Is Required for Single-Stranded DNA Binding.
L. E. Mechanic, M. E. Latta, and S. W. Matson (1999)
J. Bacteriol. 181, 2519-2526
   Abstract »    Full Text »
A Mutation in the C-terminal Putative Zn2+ Finger Motif of UL52 Severely Affects the Biochemical Activities of the HSV-1 Helicase-Primase Subcomplex.
N. Biswas and S. K. Weller (1999)
J. Biol. Chem. 274, 8068-8076
   Abstract »    Full Text »    PDF »
Equilibrium Binding Studies of Non-claret Disjunctional Protein (Ncd) Reveal Cooperative Interactions between the Motor Domains.
K. A. Foster, J. J. Correia, and S. P. Gilbert (1998)
J. Biol. Chem. 273, 35307-35318
   Abstract »    Full Text »    PDF »
Bacteriophage T7 DNA Helicase Binds dTTP, Forms Hexamers, and Binds DNA in the Absence of Mg2+. THE PRESENCE OF dTTP IS SUFFICIENT FOR HEXAMER FORMATION AND DNA BINDING.
K. M. Picha and S. S. Patel (1998)
J. Biol. Chem. 273, 27315-27319
   Abstract »    Full Text »    PDF »
Mutation of a Highly Conserved Arginine in Motif IV of Escherichia coli DNA Helicase II Results in an ATP-binding Defect.
M. C. Hall and S. W. Matson (1997)
J. Biol. Chem. 272, 18614-18620
   Abstract »    Full Text »    PDF »
The dTTPase mechanism of T7 DNA helicase resembles the binding change mechanism of the F1-ATPase.
M. M. Hingorani, M. T. Washington, K. C. Moore, and S. S. Patel (1997)
PNAS 94, 5012-5017
   Abstract »    Full Text »    PDF »
Interactions of a Subassembly of the Herpes Simplex Virus Type 1Helicase-Primase with DNA.
S. Healy, X. You, and M. Dodson (1997)
J. Biol. Chem. 272, 3411-3415
   Abstract »    Full Text »    PDF »
Strandedness Discrimination in Peptide-Polynucleotide Complexes.
N. P. Johnson, H. Mazarguil, and A. Lopez (1996)
J. Biol. Chem. 271, 19675-19679
   Abstract »    Full Text »    PDF »
The Ordered Assembly of the phi X174-type Primosome. I. ISOLATION AND IDENTIFICATION OF INTERMEDIATE PROTEIN-DNA COMPLEXES.
J. Y. Ng and K. J. Marians (1996)
J. Biol. Chem. 271, 15642-15648
   Abstract »    Full Text »    PDF »
Nucleotide and DNA-induced Conformational Changes in the Bacteriophage T7 Gene 4 Protein.
Y. Yong and L. J. Romano (1995)
J. Biol. Chem. 270, 24509-24517
   Abstract »    Full Text »    PDF »
Bacteriophage T4 Dda Helicase Translocates in a Unidirectional Fashion on Single-stranded DNA.
K. D. Raney and S. J. Benkovic (1995)
J. Biol. Chem. 270, 22236-22242
   Abstract »    Full Text »    PDF »
Strand-Specific Recognition of a Synthetic DNA Replication Fork by the SV40 Large Tumor Antigen.
D. J. SenGupta and J. A. Borowiec (1992)
Science 256, 1656-1661
   Abstract »    PDF »
The UL5 and UL52 Subunits of the Herpes Simplex Virus Type 1 Helicase-Primase Subcomplex Exhibit a Complex Interdependence for DNA Binding.
N. Biswas and S. K. Weller (2001)
J. Biol. Chem. 276, 17610-17619
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

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