Identification of a Novel RNA Splicing Pattern as a Basis of Restricted Cell Tropism of Erythrovirus B19

• Published in: Virology (New York, N.Y.) Vol. 274; no. 2; pp. 284 - 291
• Main Authors: Brunstein, John, Söderlund-Venermo, Maria, Hedman, Klaus
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.2000
• Subjects: Alternative Splicing - genetics; Amino Acid Sequence; Base Sequence; Cell Line; Cloning, Molecular; Erythrovirus B19; Humans; Introns - genetics; leader sequence; MB-02 cells; Molecular Sequence Data; Open Reading Frames - genetics; Organ Specificity; Parvovirus B19, Human - genetics; Parvovirus B19, Human - physiology; Polymerase Chain Reaction; Protein Biosynthesis; Retinoblastoma - genetics; Ribonuclease, Pancreatic - metabolism; Ribosomes - metabolism; RNA, Messenger - analysis; RNA, Messenger - genetics; RNA, Spliced Leader - analysis; RNA, Spliced Leader - genetics; RNA, Viral - analysis; RNA, Viral - genetics; splice donor site; Templates, Genetic; Transcription, Genetic - genetics; Viral Structural Proteins - biosynthesis; Viral Structural Proteins - genetics; Virus Replication
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1006/viro.2000.0460

Prior studies on the transcription of erythrovirus B19 have identified a short leader sequence associated with all spliced viral transcripts. While some variability has been observed in the acceptor for this first intron, studies to date in both permissive and nonpermissive cell types have reported a unique splice donor site. In the semipermissive MB-02 cell line, we have found that splicing of this first intron proceeds almost exclusively via a cryptic CT donor downstream of the previously reported GT donor at nucleotide 406. The resulting messages for the viral structural proteins and 11-kDa protein are thereby made bicistronic, with the first expressible polypeptide being a 34 amino acid fusion of the NS-1 and 7.5-kDa proteins. The presence of an upstream open-reading frame on these messages is likely to block effective translation of the downstream structural protein products. We propose this as a significant mechanism in determining B19's tropism on the basis of host cell splicing machinery, and present evidence in support of this model. Additionally, this is the first report of usage of a noncanonical splice donor in B19, and to our knowledge the first report of a CT-AG splice in any system.