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Mol Cell Biol, July 1998, p. 4165-4176, Vol. 18, No. 7
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Capacity of Polyomavirus Enhancer Binding Protein 2alpha B (AML1/Cbfa2) To Stimulate Polyomavirus DNA Replication Is Related to Its Affinity for the Nuclear Matrix

Lin-Feng Chen, Kosei Ito, Yota Murakami, and Yoshiaki Ito*

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 2alpha B1 (PEBP2alpha 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 PEBP2alpha B1/AML1, was also localized in the nuclear matrix fraction and competed for nuclear matrix association with PEBP2alpha B1 and GAL4-RAD. Moreover, AML1/ETO inhibited Py DNA replication stimulated by PEBP2alpha B1 and GAL4-RAD. The inhibition was specific for replication mediated by PEBP2alpha 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.

Mol Cell Biol, July 1998, p. 4165-4176, Vol. 18, No. 7
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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