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Mol Cell Biol, July 1998, p. 4165-4176, Vol. 18, No. 7
Department of Viral Oncology, Institute for
Virus Research, Kyoto University, Shogoin, Sakyo-ku, Kyoto 606, Japan
Received 26 January 1998/Returned for modification 19 February
1998/Accepted 28 April 1998
The nuclear matrix is thought to play an important role in the DNA
replication of eukaryotic cells, although direct evidence for such a
role is still lacking. A nuclear matrix-associated transcription
factor, polyomavirus (Py) enhancer binding protein 2
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Copyright © 1998, American Society for Microbiology. All rights reserved.
The Capacity of Polyomavirus Enhancer Binding
Protein 2
B (AML1/Cbfa2) To Stimulate Polyomavirus DNA
Replication Is Related to Its Affinity for the Nuclear
Matrix
B1 (PEBP2
B1)
(AML1/Cbfa2), was found to stimulate Py replication through its cognate
binding site. The minimal replication activation domain (RAD) was
identified between amino acid (aa) 302 and aa 371 by using a fusion
protein containing the GAL4 DNA binding domain (GAL4-RAD). In addition,
the region showed affinity for the nuclear matrix and, on the basis of
competition studies, binding activity for one or more proteins involved
in the initiation of Py DNA replication. A leukemogenic chimeric
protein, AML1/ETO(MTG8), which does not contain this region of
PEBP2
B1/AML1, was also localized in the nuclear matrix fraction and
competed for nuclear matrix association with PEBP2
B1 and GAL4-RAD.
Moreover, AML1/ETO inhibited Py DNA replication stimulated by
PEBP2
B1 and GAL4-RAD. The inhibition was specific for replication
mediated by PEBP2
B1 and GAL4-RAD, and proportional to the degree of
loss of these activators from the nuclear matrix, suggesting a
requirement for nuclear matrix targeting in the stimulation of Py DNA
replication by RAD. These results are the first to suggest a molecular
link between the initiation of DNA replication and the nuclear matrix compartment.
*
Corresponding author. Mailing address: Department of
Viral Oncology, Institute for Virus Research, Kyoto University,
Shogoin, Sakyo-ku, Kyoto 606, Japan. Phone: 81-75-751-4028. Fax:
81-75-752-3232. E-mail: yito{at}virus.kyoto-u.ac.jp.
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