Large T-Antigen Double Hexamers Imaged at the Simian Virus 40 Origin of Replication

doi:10.1128/MCB.20.1.34-41.2000

This Article

	Full Text
	Full Text (PDF)
	Alert me when this article is cited
	Alert me if a correction is posted

Services

	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new issues of the journal
	Download to citation manager
	Reprints and Permissions
	Copyright Information
	Books from ASM Press
	MicrobeWorld

Citing Articles

	Citing Articles via HighWire
	Citing Articles via Google Scholar

Google Scholar

	Articles by Valle, M.
	Articles by Donate, L. E.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Valle, M.
	Articles by Donate, L. E.

Previous Article | Next Article

Molecular and Cellular Biology, January 2000, p. 34-41, Vol. 20, No. 1
0270-7306/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Large T-Antigen Double Hexamers Imaged at the Simian Virus 40 Origin of Replication

Mikel Valle,¹ Claudia Gruss,² Lothar Halmer,² José M. Carazo,¹^,^* and Luis Enrique Donate¹

Centro Nacional de Biotecnología (CSIC), Campus de Cantoblanco, 28049 Madrid, Spain,¹ and University of Konstanz Department of Biology, 78457 Konstanz, Germany²

Received 14 June 1999/Returned for modification 2 August 1999/Accepted 28 September 1999

The initial step of simian virus 40 (SV40) DNA replication is the binding of the large tumor antigen (T-Ag) to the SV40 coreorigin. In the presence of Mg²⁺ and ATP, T-Ag forms a double-hexamer complex covering the completecore origin. By using electron microscopy and negative staining,we visualized for the first time T-Ag double hexamers bound tothe SV40 origin. Image processing of side views of these nucleoproteincomplexes revealed bilobed particles 24 nm long and 8 to 12 nmwide, which indicates that the two T-Ag hexamers are orientedhead to head. Taking into account all of the biochemical dataknown on the T-Ag-DNA interactions at the replication origin,we present a model in which the DNA passes through the inner channelof both hexamers. In addition, we describe a previously undetectedstructural domain of the T-Ag hexamer and thereby amend the previouslypublished dimensions of the T-Ag hexamer. This domain we havedetermined to be the DNA-binding domain of T-Ag.

^* Corresponding author. Mailing address: Centro Nacional de Biotecnologia (CSIC), Campus de Cantoblanco, 28049 Madrid, Spain.Phone: 34-91-5854543. Fax: 34-91-5854506. E-mail: carazo{at}cnb.uam.es.

Molecular and Cellular Biology, January 2000, p. 34-41, Vol. 20, No. 1
0270-7306/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

Khopde, S., Simmons, D. T. (2008). Simian Virus 40 DNA Replication Is Dependent on an Interaction between Topoisomerase I and the C-Terminal End of T Antigen. J. Virol. 82: 1136-1145 [Abstract] [Full Text]
Kumar, A., Meinke, G., Reese, D. K., Moine, S., Phelan, P. J., Fradet-Turcotte, A., Archambault, J., Bohm, A., Bullock, P. A. (2007). Model for T-Antigen-Dependent Melting of the Simian Virus 40 Core Origin Based on Studies of the Interaction of the Beta-Hairpin with DNA. J. Virol. 81: 4808-4818 [Abstract] [Full Text]
Reese, D. K., Meinke, G., Kumar, A., Moine, S., Chen, K., Sudmeier, J. L., Bachovchin, W., Bohm, A., Bullock, P. A. (2006). Analyses of the Interaction between the Origin Binding Domain from Simian Virus 40 T Antigen and Single-Stranded DNA Provide Insights into DNA Unwinding and Initiation of DNA Replication. J. Virol. 80: 12248-12259 [Abstract] [Full Text]
Duggin, I. G., Bell, S. D. (2006). The Chromosome Replication Machinery of the Archaeon Sulfolobus solfataricus. J. Biol. Chem. 281: 15029-15032 [Abstract] [Full Text]
Meinke, G., Bullock, P. A., Bohm, A. (2006). Crystal Structure of the Simian Virus 40 Large T-Antigen Origin-Binding Domain. J. Virol. 80: 4304-4312 [Abstract] [Full Text]
Gomez-Llorente, Y., Fletcher, R. J., Chen, X. S., Carazo, J. M., Martin, C. S. (2005). Polymorphism and Double Hexamer Structure in the Archaeal Minichromosome Maintenance (MCM) Helicase from Methanobacterium thermoautotrophicum. J. Biol. Chem. 280: 40909-40915 [Abstract] [Full Text]
Weisshart, K., Friedl, S., Taneja, P., Nasheuer, H.-P., Schlott, B., Grosse, F., Fanning, E. (2004). Partial Proteolysis of Simian Virus 40 T Antigen Reveals Intramolecular Contacts between Domains and Conformation Changes upon Hexamer Assembly. J. Biol. Chem. 279: 38943-38951 [Abstract] [Full Text]
Gai, D., Li, D., Finkielstein, C. V., Ott, R. D., Taneja, P., Fanning, E., Chen, X. S. (2004). Insights into the Oligomeric States, Conformational Changes, and Helicase Activities of SV40 Large Tumor Antigen. J. Biol. Chem. 279: 38952-38959 [Abstract] [Full Text]
Sclafani, Robert. A., Fletcher, R. J., Chen, X. S. (2004). Two heads are better than one: regulation of DNA replication by hexameric helicases. Genes Dev. 18: 2039-2045 [Full Text]
Reese, D. K., Sreekumar, K. R., Bullock, P. A. (2004). Interactions Required for Binding of Simian Virus 40 T Antigen to the Viral Origin and Molecular Modeling of Initial Assembly Events. J. Virol. 78: 2921-2934 [Abstract] [Full Text]
Jiao, J., Simmons, D. T. (2003). Nonspecific Double-Stranded DNA Binding Activity of Simian Virus 40 Large T Antigen Is Involved in Melting and Unwinding of the Origin. J. Virol. 77: 12720-12728 [Abstract] [Full Text]
Roy, R., Trowbridge, P., Yang, Z., Champoux, J. J., Simmons, D. T. (2003). The Cap Region of Topoisomerase I Binds to Sites near Both Ends of Simian Virus 40 T Antigen. J. Virol. 77: 9809-9816 [Abstract] [Full Text]
Makhov, A. M., Lee, S. S.-K., Lehman, I. R., Griffith, J. D. (2003). Origin-specific unwinding of herpes simplex virus 1 DNA by the viral UL9 and ICP8 proteins: Visualization of a specific preunwinding complex. Proc. Natl. Acad. Sci. USA 100: 898-903 [Abstract] [Full Text]
Alexandrov, A. I., Botchan, M. R., Cozzarelli, N. R. (2002). Characterization of Simian Virus 40 T-antigen Double Hexamers Bound to a Replication Fork. THE ACTIVE FORM OF THE HELICASE. J. Biol. Chem. 277: 44886-44897 [Abstract] [Full Text]
Kim, R. J., Moine, S., Reese, D. K., Bullock, P. A. (2002). Peptides Containing Cyclin/Cdk-Nuclear Localization Signal Motifs Derived from Viral Initiator Proteins Bind to DNA When Unphosphorylated. J. Virol. 76: 11785-11792 [Abstract] [Full Text]
Ott, R. D., Wang, Y., Fanning, E. (2002). Mutational Analysis of Simian Virus 40 T-Antigen Primosome Activities in Viral DNA Replication. J. Virol. 76: 5121-5130 [Abstract] [Full Text]
Purviance, J. D., Prack, A. E., Barbaro, B., Bullock, P. A. (2001). In the Simian Virus 40 In Vitro Replication System, Start Site Selection by the Polymerase {alpha}-Primase Complex Is Not Significantly Altered by Changes in the Concentration of Ribonucleotides. J. Virol. 75: 6392-6401 [Abstract] [Full Text]
Wu, C., Roy, R., Simmons, D. T. (2001). Role of Single-Stranded DNA Binding Activity of T Antigen in Simian Virus 40 DNA Replication. J. Virol. 75: 2839-2847 [Abstract] [Full Text]
Tang, Q., Bell, P., Tegtmeyer, P., Maul, G. G. (2000). Replication but Not Transcription of Simian Virus 40 DNA Is Dependent on Nuclear Domain 10. J. Virol. 74: 9694-9700 [Abstract] [Full Text]
Sreekumar, K. R., Prack, A. E., Winters, D. R., Barbaro, B. A., Bullock, P. A. (2000). The Simian Virus 40 Core Origin Contains Two Separate Sequence Modules That Support T-Antigen Double-Hexamer Assembly. J. Virol. 74: 8589-8600 [Abstract] [Full Text]
Barbaro, B. A., Sreekumar, K. R., Winters, D. R., Prack, A. E., Bullock, P. A. (2000). Phosphorylation of Simian Virus 40 T Antigen on Thr 124 Selectively Promotes Double-Hexamer Formation on Subfragments of the Viral Core Origin. J. Virol. 74: 8601-8613 [Abstract] [Full Text]
Gai, D., Roy, R., Wu, C., Simmons, D. T. (2000). Topoisomerase I Associates Specifically with Simian Virus 40 Large-T-Antigen Double Hexamer-Origin Complexes. J. Virol. 74: 5224-5232 [Abstract] [Full Text]
White, P. W., Pelletier, A., Brault, K., Titolo, S., Welchner, E., Thauvette, L., Fazekas, M., Cordingley, M. G., Archambault, J. (2001). Characterization of Recombinant HPV6 and 11 E1 Helicases. EFFECT OF ATP ON THE INTERACTION OF E1 WITH E2 AND MAPPING OF A MINIMAL HELICASE DOMAIN. J. Biol. Chem. 276: 22426-22438 [Abstract] [Full Text]
Tye, B. K., Sawyer, S. (2000). The Hexameric Eukaryotic MCM Helicase: Building Symmetry from Nonidentical Parts. J. Biol. Chem. 275: 34833-34836 [Full Text]

J. Bacteriol.	J. Virol.	Eukaryot. Cell

Microbiol. Mol. Biol. Rev.	Clin. Vaccine Immunol.	All ASM Journals