Interaction of a nuclear factor-1-like protein with the regulatory region of the human polyomavirus JC virus.
We have initiated a study to identify host proteins which interact with the regulatory region of the human polyomavirus JC (JCV), which is associated with the demyelinating disease, progressive multifocal leukoencephalopathy. We examined the interaction of nuclear proteins prepared from different cell lines with the JCV regulatory region by DNA binding gel retardation assays. Binding was detected with nuclear extracts prepared from human fetal glial cells, glioma cells, and HeLa cells. Little or no binding was detected with nuclear extracts prepared from human embryonic kidney cells. Competitive binding assays suggest that the nuclear factor(s) which interacted with the JCV regulatory region was different from those which interacted with the regulatory region of the closely related polyomavirus SV40. We found three areas in the JCV regulatory region protected from DNase I digestion: site A, located just upstream from the TATA sequence in the first 98-base pair (bp) repeat; site B, located upstream from the TATA sequence in the second 98-bp repeat; and site C, located just following the second 98-bp repeat. There were some differences in the ability of the nuclear factor(s) from the two brain cell lines and HeLa cells to completely protect the nucleotides within the footprint region. The results from the DNase I protective studies and competitive DNA binding studies with specific oligonucleotides, suggest that nuclear factor-1 or a nuclear factor-1-like factor is interacting with all three sites in the JCV regulatory region. In addition, the results suggest that the nuclear factor which interacts with the JCV regulatory region from human brain cell lines is different from the factor found in HeLa cells.
Amemiya
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K
Infectious Diseases Branch, National Institute of Neurological and Communicative Disorders and Stroke, Bethesda, Maryland 20892.
Traub
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R
Durham
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Major
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D001483:Base Sequence; D001667:Binding, Competitive; D022762:CCAAT-Enhancer-Binding Proteins; D002460:Cell Line; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000235:genetics; D004268:DNA-Binding Proteins / Q000502:physiology*; D003850:Deoxyribonuclease I / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D006801:Humans; D066298:In Vitro Techniques; D007577:JC Virus / Q000235:genetics*; D051839:NFI Transcription Factors; D009687:Nuclear Proteins / Q000502:physiology*; D011120:Polyomavirus / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000502:physiology*; D051840:Y-Box-Binding Protein 1
D022762:CCAAT-Enhancer-Binding Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D051839:NFI Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors; D051840:Y-Box-Binding Protein 1; C070856:YBX1 protein, human; D003850:Deoxyribonuclease I