Defibrinated bovine plasma inhibits retroviral transcription by blocking p52 activation of the NFkappaB element in the long terminal repeat.
Bovine leukemia virus (BLV) induces a persistent but latent infection in cattle. Viral latency is invoked by a protein known as plasma blocking factor (PBF) that is found in both bovine and human plasma. We report here on pathways that mediate latency in the presence of PBF. Reporter-gene constructs driven by the promoters of 6 retroviruses were used to measure the production of chloramphenicol acetyl transferase (CAT) in cell lines cultured with or without defibrinated bovine plasma. Plasma inhibited CAT production only in constructs containing an NFkappaB-binding element proximal to the initiation site (BLV, human immunodeficiency virus, and human T-cell leukemia virus). The promoters of Bovine immunodeficiency virus, Feline immunodeficiency virus, or Feline leukemia virus were not inhibited in the presence of bovine plasma. Using gel mobility shift assays, we demonstrated that activation of viral transcription upon stimulation with phorbol esters and ionomycin was mediated through the NFkappaB element and that this was abrogated in the presence of plasma. Furthermore, analysis of individual NFkappaB proteins in nuclear extracts of mononuclear cells or Jurkat cells showed that all 5 members of the NFkappaB family were upregulated in response to stimulation, but only p52 was significantly downregulated in the presence of bovine plasma. Thus, we infer that plasma effects are mediated through interference with either p52 translocation to the nucleus or p52 synthesis.
van den Heuvel
Marianne J
MJ
Department of Pediatrics, University of Western Ontario, London, Ontario. mvandenh@uwo.ca
Copeland
Karen F
KF
Cates
Elizabeth C
EC
Jefferson
Barbara J
BJ
Jacobs
Robert M
RM
D000818:Animals; D000951:Antigens, Neoplasm / Q000096:biosynthesis*; D002417:Cattle; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis*; D005260:Female; D015967:Gene Expression Regulation, Viral; D001909:Leukemia Virus, Bovine / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D016328:NF-kappa B; D010949:Plasma; D020079:Terminal Repeat Sequences; D014158:Transcription, Genetic; D014162:Transfection; D017735:Virus Latency
D000951:Antigens, Neoplasm; D016328:NF-kappa B; C020163:blocking factor; D015500:Chloramphenicol O-Acetyltransferase