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 core origin. In
the presence of Mg2+ and ATP, T-Ag forms a double-hexamer
complex covering the complete core origin. By using electron microscopy
and negative staining, we visualized for the first time T-Ag double
hexamers bound to the SV40 origin. Image processing of side views of
these nucleoprotein complexes revealed bilobed particles 24 nm long and
8 to 12 nm wide, which indicates that the two T-Ag hexamers are
oriented head to head. Taking into account all of the biochemical data known on the T-Ag-DNA interactions at the replication origin, we
present a model in which the DNA passes through the inner channel of
both hexamers. In addition, we describe a previously undetected structural domain of the T-Ag hexamer and thereby amend the previously published dimensions of the T-Ag hexamer. This domain we have determined 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]