title: Interactions between c-Jun, nuclear factor 1, and JC virus promoter sequences: implications for viral tropism.
abstract: The infectious cycle of the human polyomavirus JC (JCV) is ultimately regulated in cellular nuclei at the level of viral protein expression and genomic replication. Such activity is prompted by interactions between variant nucleotide sequences within the JCV regulatory region (promoter) and cellular transcription factors that bind specific DNA consensus sites. In previous work we identified an NF-1 class member, NF-1X, as a critical transcription factor affecting the JCV cellular host range. Within variant JCV promoters, as well as other viral and cellular promoters, adjacently located NF-1 and AP-1 consensus sites are often found. The close proximity of these two binding sites suggests the opportunity for interaction between NF-1 and AP-1 proteins. Here, by electrophoretic mobility shift assays, we show temporal and dose-dependent interference by an AP-1 family member, c-Jun, upon NF-1 proteins binding an NF-1 consensus site derived from JCV promoter sequence. Moreover, as demonstrated by protein-protein interaction assays, we identify specific binding affinity independent of DNA binding between NF-1X and c-Jun. Finally, to compare the binding profiles of NF-1X and c-Jun on JCV promoter sequence in parallel with in vivo detection of viral activity levels, we developed an anchored transcriptional promoter (ATP) assay. With use of extracts from JCV-infected cells transfected to overexpress either NF-1X or c-Jun, ATP assays showed concurrent increases in NF-1X binding and viral protein expression. Conversely, increased c-Jun binding accompanied decreases in both NF-1X binding and viral protein expression. Therefore, inhibition of NF-1X binding by c-Jun appears to play a role in regulating levels of JCV activity.
authors.affiliation: Laboratory of Molecular Medicine and Neuroscience, NINDS, NIH, 10 Center Drive, Building 10, Room 3B14, MSC1296, Bethesda, Maryland 20892-1296, USA.
authors.forename: Veerasamy
authors.initials: V
authors.lastname: Ravichandran
authors.forename: Bruce F
authors.initials: BF
authors.lastname: Sabath
authors.forename: Peter N
authors.initials: PN
authors.lastname: Jensen
authors.forename: Sidney A
authors.initials: SA
authors.lastname: Houff
authors.forename: Eugene O
authors.initials: EO
authors.lastname: Major
mesh_terms: D001665:Binding Sites / Q000235:genetics; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D005814:Genes, Viral; D006801:Humans; D066298:In Vitro Techniques; D007577:JC Virus / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D048031:JNK Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D039902:Multipotent Stem Cells / Q000821:virology; D051839:NFI Transcription Factors / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000821:virology; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004279:DNA, Viral; D051839:NFI Transcription Factors; D011993:Recombinant Fusion Proteins; D048031:JNK Mitogen-Activated Protein Kinases