AEGiS-11IAC: Possible regulation of HIV replication by a cellular factor binding to the primer binding site.

11th International AIDS Conference


Vancouver, British Columbia — July 7-12, 1996

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Possible regulation of HIV replication by a cellular factor binding to the primer binding site.

Int Conf AIDS 1996 Jul 7-12; 11:58 (abstract no. Mo.A.1013)
Lai D, Guo HG, Gallo RC, Reitz M; UMBI, Baltimore, Maryland. Fax: 410-706-8184.

OBJECTIVE: The regulation of HIV replication has been intensively studied. One of the early steps is the reverse transcription of viral RNA into viral DNA, which is then integrated into the infected host cell genome. The natural primers for reverse transcription are the cell derived tRNAs. However, the exact interactions of the primer binding site (PBS) with the viral RNA, the tRNA primer, HIV reverse transcriptase and cellular factor(s) are still unclear. The present study is intended to help understand these interactions by characterizing a possible cellular protein which binds to the PBS on viral RNA.

METHODS: The primer binding site (PBS) of HIV-1 is an 18 nucleotide sequence immediately 3' to the left hand LTR which complements the 3' end of tRNA1ys3. The phosphorothio-oligonucleotide 24mer (lys3) and different control oligomer were added to PM-1 cells acutely infected with HIV-1(Bal) or SuptI cells acutely infected with HIV-2. Viral expression was monitored by p24 or p27 antigen capture assays. Gel retardation assays were performed with this oligonucleotide, which was also used as a probe to screen a human lymphocyte cDNA expression library.

RESULTS: The PBS oligonucleotide induced HIV-1Bal and HIV-2 expression at least 8 fold in vitro. A non-specific induction of cytokines mediated by a reaction to non-methylated CpG pairs was ruled out by use of methylated oligomers. We hypothesize that this oligonucleotide binds to a cellular factor(s) involved in regulation of the HIV primer binding step, causes depletion of the cellular factor(s), and thus induces HIV replication. The gel retardation assays have shown specific binding of cellular protein(s) with this oligonucleotide. Specific clones have been selected from human library.

CONCLUSIONS: This work may provide evidence for cellular control of early stages of HIV replication.

Keywords: AEGIS, Virus Replication, Binding Sites, RNA, Viral, HIV-1, HIV-2, RNA-Directed DNA Polymerase, RNA, Transfer, DNA, Viral, DNA Primers, Transcription, Genetic, Human, In Vitro, pharmacokinetics, virology, genetics, ICA11KWDaegis,virusreplication,bindingsites,rna,viral,hiv-1,hiv-2,rna-directeddnapolymerase,rna,transfer,dna,viral,dnaprimers,transcription,genetic,human,invitro,pharmacokinetics,virology,genetics,ica11

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MoA1013

Copyright © 1996 - International AIDS Society (IAS). Reproduction of this abstract (other than one copy for personal reference) must be cleared through the IAS.