title: Rearranged JC virus noncoding control regions found in progressive multifocal leukoencephalopathy patient samples increase virus early gene expression and replication rate.
abstract: Polyomavirus JC (JCV) infects ∼ 60% of the general population, followed by asymptomatic urinary shedding in ∼ 20%. In patients with pronounced immunodeficiency, including HIV/AIDS, JCV can cause progressive multifocal leukoencephalopathy (PML), a devastating brain disease of high mortality. While JCV in the urine of healthy people has a linear noncoding control region called the archetype NCCR (at-NCCR), JCV in brain and cerebrospinal fluid (CSF) of PML patients bear rearranged NCCRs (rr-NCCRs). Although JCV NCCR rearrangements are deemed pathognomonic for PML, their role as a viral determinant is unclear. We sequenced JCV NCCRs found in CSF of eight HIV/AIDS patients newly diagnosed with PML and analyzed their effect on early and late gene expression using a bidirectional reporter vector recapitulating the circular polyomavirus early and late gene organization. The rr-NCCR sequences were highly diverse, but all increased viral early reporter gene expression in progenitor-derived astrocytes, glia-derived cells, and human kidney compared to the expression levels with the at-NCCR. The expression of simian virus 40 (SV40) large T antigen or HIV Tat expression in trans was associated with a strong increase of at-NCCR-controlled early gene expression, while rr-NCCRs were less responsive. The insertion of rr-NCCRs into the JCV genome backbone revealed higher viral replication rates for rr-NCCR compared to those of the at-NCCR JCV in human progenitor-derived astrocytes or glia cells, which was abrogated in SV40 large T-expressing COS-7 cells. We conclude that naturally occurring JCV rr-NCCR variants from PML patients confer increased early gene expression and higher replication rates compared to those of at-NCCR JCV and thereby increase cytopathology.
authors.affiliation: Transplantation Virology, Institute for Medical Microbiology, Department of Biomedicine, University of Basel, CH-4003 Basel, Switzerland.
authors.forename: Rainer
authors.initials: R
authors.lastname: Gosert
authors.forename: Piotr
authors.initials: P
authors.lastname: Kardas
authors.forename: Eugene O
authors.initials: EO
authors.lastname: Major
authors.forename: Hans H
authors.initials: HH
authors.lastname: Hirsch
mesh_terms: D000328:Adult; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000134:cerebrospinal fluid / Q000235:genetics / Q000652:urine; D005260:Female; D015870:Gene Expression; D015321:Gene Rearrangement; D017930:Genes, Reporter; D005814:Genes, Viral; D006801:Humans; D007577:JC Virus / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity / Q000502:physiology*; D007968:Leukoencephalopathy, Progressive Multifocal / Q000821:virology*; D008297:Male; D008875:Middle Aged; D014162:Transfection; D014779:Virus Replication
chemical_list: D017931:DNA Primers; D004279:DNA, Viral