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The effects of measles virus persistent infection on AP-1 transcription factor binding in neuroblastoma cells
D Fishman1, M Wolfson, E Bazarski
1Department of Microbiology and Immunology, Faculty of Health Sciences, The Cancer Research Center, Ben Gurion University of the Negev, Beer Sheva, Israel.
Abstract:
Measles virus (MV) persistence in brain cells has broad effects on different cellular functions. We have previously shown that NS20Y clone, originally derived from C1300 neuroblastoma cells, persistently infected with MV (NS20Y/MS), displays constitutively elevated levels of c-fos and PKC mRNAs, implying MV-mediated effects on transcriptional regulation. Nonetheless, the mode by which virus affects the transcriptional machinery still remains obscure. In order to define this phenomenon, we studied the binding properties of major transcription factors (AP-1 and NFkappaB) in NS20Y/MS cells. Using electrophoretic mobility shift approach (EMSA) with the appropriate oligonucleotide probes, we have found that the persistent MV infection does not affect NFkappaB binding, while the AP-1 binding was significantly decreased. Similar inhibition was not observed in NS20Y cells acutely infected with MV. Anti-measles antibody-mediated restriction of viral gene expression restored AP-1 binding, thus suggesting that measles virus proteins may affect the components of the host transcriptional machinery.
Insights
Persistent measles virus (MV) infection in brain cells decreases AP-1 transcription factor binding. Measles virus proteins, not acute infection, appear to alter host transcriptional machinery, impacting cellular functions.
Area of Science:
- Neurovirology
- Molecular Biology
- Cellular Neuroscience
Background:
- Measles virus (MV) persistence in brain cells affects cellular functions, including transcriptional regulation.
- Elevated c-fos and PKC mRNAs in persistently infected neuroblastoma cells (NS20Y/MS) suggest MV impacts transcriptional regulation.
- The precise mechanism by which MV influences the host transcriptional machinery remains unclear.
Purpose of the Study:
- To investigate the impact of persistent measles virus infection on the binding activity of key transcription factors, AP-1 and NF-kappaB, in neuroblastoma cells.
- To differentiate the effects of persistent versus acute MV infection on transcription factor binding.
- To explore the role of measles virus proteins in modulating host transcriptional machinery.
Main Methods:
- Electrophoretic mobility shift assay (EMSA) was employed to assess the DNA-binding activity of AP-1 and NF-kappaB.
- Oligonucleotide probes specific for AP-1 and NF-kappaB binding sites were utilized.
- Experiments were conducted on persistently infected (NS20Y/MS) and acutely infected (NS20Y) neuroblastoma cells.
- The effect of anti-measles antibody treatment on viral gene expression and AP-1 binding was examined.
Main Results:
- Persistent measles virus infection in NS20Y/MS cells significantly decreased the binding activity of the AP-1 transcription factor.
- NF-kappaB binding activity remained unaffected by persistent MV infection.
- Acute measles virus infection of NS20Y cells did not result in the observed inhibition of AP-1 binding.
- Restoration of AP-1 binding activity was observed upon anti-measles antibody-mediated restriction of viral gene expression in persistently infected cells.
Conclusions:
- Persistent measles virus infection, but not acute infection, leads to a significant reduction in AP-1 transcription factor binding in neuroblastoma cells.
- Measles virus proteins are implicated in the observed modulation of host transcriptional machinery, specifically affecting AP-1 binding.
- These findings suggest a mechanism by which measles virus persistence disrupts cellular functions through alterations in host gene regulation.