1. Limitation of Tat-Associated Transcriptional Processivity in HIV-Infected PBMC
- Author
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Melanie Adams, Joseph M Romeo, Christine Wong, and Dan Wang
- Subjects
Gene Expression Regulation, Viral ,Transcription, Genetic ,T-Lymphocytes ,T cell ,Biology ,Peripheral blood mononuclear cell ,Cell Line ,03 medical and health sciences ,0302 clinical medicine ,Transcription (biology) ,In vivo ,Virology ,medicine ,Humans ,RNA, Messenger ,RNA Processing, Post-Transcriptional ,030304 developmental biology ,0303 health sciences ,Messenger RNA ,RNA ,U937 Cells ,Processivity ,Molecular biology ,Phenotype ,3. Good health ,medicine.anatomical_structure ,Gene Products, tat ,HIV-1 ,Leukocytes, Mononuclear ,RNA, Viral ,tat Gene Products, Human Immunodeficiency Virus ,Mitogens ,030217 neurology & neurosurgery - Abstract
The ability of HIV to match levels of viral mRNA to the activation state of the host cell may play a role in its ability to persist as well as to replicate. This linkage depends on the function of the viral transcriptional regulatory protein, Tat, which increases the efficiency of RNA elongation (transcriptional processivity) in response to cellular activation. To quantify levels of Tat functionin vivo,a quantitative competitive RT-PCR assay was developed that reflects levels of TAR leader fragments (nonprocessive transcripts) and viral mRNA (processive transcripts), indicating low or high levels of Tat function, respectively. The abundance of these RNA species was measured in peripheral blood mononuclear cells (PBMC) of 22 HIV-1-positive individuals (CD4+T cell counts 63–934/mm3) and in established cell line models of HIV constitutive replication (H9IIIB) and reversible latency (U1 and ACH-2). In PBMC, the level of total viral transcripts ranged over four orders of magnitude; however, nonprocessive transcription predominated: 70% of PBMC samples had a ratio of processive to total transcripts of
- Published
- 1999