title: Homologous SV40 RNA trans-splicing: a new mechanism for diversification of viral sequences and phenotypes.
abstract: Simian Virus 40 (SV40) is a polyomavirus found in both monkeys and humans, which causes cancer in some animal models. In humans, SV40 has been reported to be associated with cancers but causality has not been proven yet. The transforming activity of SV40 is mainly due to its 94-kD large T antigen, which binds to the retinoblastoma (pRb) and p53 tumor suppressor proteins, and thereby perturbs their functions. Here we describe a 100 kD super T antigen harboring a duplication of the pRB binding domain that was associated with unusual high cell transformation activity and that was generated by a novel mechanism involving homologous RNA trans-splicing of SV40 early transcripts in transformed rodent cells. Enhanced trans-splice activity was observed in clones carrying a single point mutation in the large T antigen 5' donor splice site (ss). This mutation impaired cis-splicing in favor of an alternative trans-splice reaction via a cryptic 5'ss within a second cis-spliced SV40 pre-mRNA molecule and enabled detectable gene expression. Next to the cryptic 5'ss we identified additional trans-splice helper functions, including putative dimerization domains and a splice enhancer sequence. Our findings suggest RNA trans-splicing as a SV40-intrinsic mechanism that supports the diversification of viral RNA and phenotypes.
authors.affiliation: Institut fuer Molekularbiologie und Biochemie; Freie Universität Berlin; Berlin, German.
authors.forename: Joachim
authors.initials: J
authors.lastname: Eul
authors.affiliation: Department of Microbiology; Yong Loo Lin School of Medicine; National University of Singapore; Singapore.
authors.forename: Volker
authors.initials: V
authors.lastname: Patzel
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D005091:Exons; D010641:Phenotype; D022821:RNA Splice Sites / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D051381:Rats; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D020040:Trans-Splicing
chemical_list: D000957:Antigens, Viral, Tumor; D022821:RNA Splice Sites; D012367:RNA, Viral
keywords: SV40 100 kD super T antigen; alternative splicing; cell transformation; molecular basis of diseases; trans-splicing; viral gene expression