Expression and characterization of the Borna disease virus polymerase.
Borna disease virus is the prototype of a new family, Bornaviridae, within the order Mononegavirales, that is characterized by nuclear transcription, splicing, low level replication, and neurotropism. The products of five open reading frames predicted from the genomic sequence have been confirmed; however, expression of the sixth, corresponding to the putative viral polymerase (L), has not been demonstrated. Here, we describe expression and characterization of a 190-kDa protein proposed to represent L. Expression of this protein from the third transcription unit of the viral genome is dependent on a splicing event that fuses a small upstream open reading frame in frame with the larger downstream continuous open reading frame. The protein is detected by serum antibodies from infected rats and is present in the nucleus, where it colocalizes with the phosphoprotein. L is also shown to be phosphorylated by cellular kinases and to interact with the viral phosphoprotein in coimmunoprecipitation studies. These findings are consistent with the identity of the 190-kDa protein as the viral polymerase and provide insights and describe reagents that will be useful for Bornavirus molecular biology and pathobiology.
Walker
M P
MP
Emerging Diseases Laboratory, Departments of Neurology, Anatomy and Neurobiology, and Microbiology and Molecular Genetics, University of California, Irvine, California 92697-4292, USA.
Jordan
I
I
Briese
T
T
Fischer
N
N
Lipkin
W I
WI
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001890:Borna Disease / Q000276:immunology; D001891:Borna disease virus / Q000201:enzymology*; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D010766:Phosphorylation; D012326:RNA Splicing; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D011917:Rats, Inbred Lew; D014162:Transfection
D000914:Antibodies, Viral; D012324:RNA-Dependent RNA Polymerase