Authentic Borna disease virus transcripts are spliced less efficiently than cDNA-derived viral RNAs.
Borna disease virus (BDV) is a non-segmented, negative-strand RNA virus that replicates and transcribes its genome in the nucleus of infected cells. It uses the cellular splicing machinery to generate a set of alternatively spliced mRNAs from the 2.8 and 7.1 kb primary transcripts, each harbouring two introns. To determine whether splicing of these transcripts is regulated by viral factors, the extent of splicing was studied in infected cells and COS-7 cells transiently transfected with plasmids encoding the 2.8 kb RNA of BDV. Unspliced RNA was found to be the most abundant RNA species in infected cells, whereas viral transcripts lacking both introns were only found in minute amounts. In sharp contrast, plasmid-derived 2.8 kb RNA was predominantly intron 1-spliced and double-spliced. Co-expression of the BDV proteins P, N and X did not influence splicing of plasmid-expressed 2.8 kb RNA. Furthermore, the splicing pattern did not change when the 2.8 kb RNA was expressed in BDV-infected cells. Based on these results we speculate that splicing of authentic BDV transcripts is tightly linked to transcription by the viral polymerase.
Jehle
Christian
C
Department of Virologie, Institute for Medical Microbiology & Hygiene, University of Freiburg, Hermann-Herder Str. 11, D-79104 Freiburg, Germany1.
Lipkin
W Ian
WI
Emerging Diseases Laboratory, Departments of Neurology, Anatomy and Neurobiology, and Microbiology and Molecular Genetics, University of California-Irvine, Irvine, CA 92697-4292, USA2.
Staeheli
Peter
P
Department of Virologie, Institute for Medical Microbiology & Hygiene, University of Freiburg, Hermann-Herder Str. 11, D-79104 Freiburg, Germany1.
Marion
Rosa M
RM
Centro Nacional de Biotecnologia (CSIC), Cantoblanco, 28049 Madrid, Spain3.
Schwemmle
Martin
M
Department of Virologie, Institute for Medical Microbiology & Hygiene, University of Freiburg, Hermann-Herder Str. 11, D-79104 Freiburg, Germany1.
D000818:Animals; D001891:Borna disease virus / Q000235:genetics*; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D010957:Plasmids; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis*
D018076:DNA, Complementary; D012333:RNA, Messenger; D012367:RNA, Viral