A high throughput RNAi screen reveals determinants of HIV-1 activity in host kinases.

Abstract

Drug resistance remains a great challenge in HIV/AIDS treatment despite the recent advances in novel therapeutics. It may be a good strategy to develop drugs targeting the essential host factors to decrease the risk of drug resistance. Previous studies suggested that so many host kinases play roles in HIV life cycles. More importantly, many kinase genes are drugable targets, therefore, it is vital to figure out host kinases responsible for HIV-1 infection and replication to provide novel therapeutic regimens and to deepen our understanding to HIV-host interaction. In present work, a high throughput RNAi screen with a shRNA library against 474 kinases was applied to HEK293T cells stably expressed a HIV-1 LTR (long terminal repeat)-driven reporter plasmid. Four genes, AK1, EphB2, PRKACB and CDK5R2, were found to specifically suppress the HIV-1 LTR activity following effective knockdown. Furthermore, overexpression of AK1 and PRKACB upregulated the HIV-1 LTR activity. Therefore, AK1 and PRKACB are in positive control of HIV-1 activity. DNA microarray analysis demonstrated that overlapped genes between AK1-silenced and PRKACB-silenced cells were mainly enriched in several amino acid biosynthesis pathways, TGF-β signaling and p53 signaling pathways. These alterations may repress the viral infection by the downregulation of ERK1/2, p38MAPK and NFκB signaling pathways. Collectively, our work uncovers several host kinases involving the HIV-1 infection and may provide potential therapeutic targets for AIDS treatment in future.

• Authors: Wei-Min Jiang, Xin-Yun Zhang, Yun-Zhi Zhang, Li Liu, Hong-Zhou Lu
• Journal: International journal of clinical and experimental pathology (Int J Clin Exp Pathol) Vol. 7 Issue 5 Pg. 2229-37 ( 2014) ISSN: 1936-2625 [Electronic] United States
• PMID: 24966931 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• RNA, Small Interfering
• Protein Kinases
• EPHB2 protein, human
• Receptor, EphB2
• Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
• PRKACB protein, human
• Adenylate Kinase
• adenylate kinase 1

Topics

• Adenylate Kinase (genetics, metabolism)
• Cyclic AMP-Dependent Protein Kinase Catalytic Subunits (genetics, metabolism)
• Gene Expression Regulation, Viral
• Gene Regulatory Networks
• HEK293 Cells
• HIV Long Terminal Repeat (genetics)
• HIV-1 (genetics, growth & development)
• High-Throughput Nucleotide Sequencing (methods)
• Host-Pathogen Interactions
• Humans
• Oligonucleotide Array Sequence Analysis
• Protein Interaction Maps
• Protein Kinases (genetics, metabolism)
• RNA Interference
• RNA, Small Interfering (genetics, metabolism)
• Receptor, EphB2 (genetics, metabolism)
• Signal Transduction
• Transfection
• Virus Replication