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Journal of Virology, September 1999, p. 7543-7555, Vol. 73, No. 9
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Sequence Requirements for the Assembly of Simian
Virus 40 T Antigen and the T-Antigen Origin Binding Domain on the
Viral Core Origin of Replication
Henry Y.
Kim,
Brett A.
Barbaro,
Woo S.
Joo,
Andrea E.
Prack,
K. R.
Sreekumar, and
Peter A.
Bullock*
Department of Biochemistry, Tufts University School
of Medicine, Boston, Massachusetts
Received 19 April 1999/Accepted 3 June 1999
The regions of the simian virus 40 (SV40) core origin that are
required for stable assembly of virally encoded T antigen (T-ag) and
the T-ag origin binding domain (T-ag-obd131-260) have been
determined. Binding of the purified T-ag-obd131-260 is
mediated by interactions with the central region of the core origin,
site II. In contrast, T-ag binding and hexamer assembly requires a
larger region of the core origin that includes both site II and an
additional fragment of DNA that may be positioned on either side of
site II. These studies indicate that in the context of T-ag, the origin
binding domain can engage the pentanucleotides in site II only if a
second region of T-ag interacts with one of the flanking sequences. The
requirements for T-ag double-hexamer assembly are complex; the
nucleotide cofactor present in the reaction modulates the sequence
requirements for oligomerization. Nevertheless, these experiments
provide additional evidence that only a subset of the SV40 core origin
is required for assembly of T-ag double hexamers.
*
Corresponding author. Mailing address: Department of
Biochemistry A703, Tufts University School of Medicine, 136 Harrison Ave., Boston, MA 02111. Phone: (617) 636-6874. Fax: (617) 636-2409. E-mail: PBULLOCK{at}OPAL.TUFTS.EDU.

Present address: Cellular Biochemistry and Biophysics Program,
Memorial Sloan-Kettering Cancer Center, New York, NY
10021.
Journal of Virology, September 1999, p. 7543-7555, Vol. 73, No. 9
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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