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Journal of Virology, August 2000, p. 6784-6789, Vol. 74, No. 15
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Evidence for Nucleic Acid Binding Ability and
Nucleosome Association of Bombyx mori Nucleopolyhedrovirus
BRO Proteins
Evgueni A.
Zemskov,1,2
WonKyung
Kang,1,* and
Susumu
Maeda1,3,
Laboratory of Molecular Entomology and
Baculovirology1 and Laboratory of CAG
Repeat Diseases, Brain Science Institute,2
RIKEN, 2-1 Hirosawa, Wako 351-0198, and Core Research for
Evolutional Science and Technology Project, Japan Science and
Technology Corporation, Kawaguchi,3 Japan
Received 22 February 2000/Accepted 2 May 2000
The Bombyx mori nucleopolyhedrovirus (BmNPV) genome
contains five related members of the bro gene family, all
of which are actively expressed in infected BmN cells. Although their
functions are unknown, their amino acid sequences contain a motif found in all known viral and prokaryotic single-stranded DNA binding proteins. To determine if they bind to nucleic acids, we fractionated the nuclei of BmNPV-infected BmN cells using a histone extraction protocol. We detected BRO-A, BRO-C, and BRO-D in the histone H1 fraction using anti-BRO antibodies. Micrococcal nuclease treatment released these BRO proteins from the chromatin fraction, suggesting their involvement in nucleosome structures. Chromatographic
fractionation showed that BRO-A and/or BRO-C interacted with core
histones. Expression of partial sequences of BRO-A proved that the
N-terminal 80 amino acid residues were required for DNA binding
activity. We also demonstrated that BmNPV BRO proteins underwent
phosphorylation and ubiquitination followed by proteasome degradation,
which may explain their distribution in the cytoplasm as well as the
nucleus. We propose that BRO-A and BRO-C may function as DNA binding
proteins that influence host DNA replication and/or transcription.
*
Corresponding author. Mailing address: Laboratory of
Molecular Entomology and Baculovirology, RIKEN, 2-1 Hirosawa, Wako
351-0198, Japan. Phone: 81-48-467-9584. Fax: 81-48-462-4678. E-mail: wkkang{at}postman.riken.go.jp.

Deceased.
Journal of Virology, August 2000, p. 6784-6789, Vol. 74, No. 15
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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