Inhibition of HIV-1 reverse transcriptase by RNA aptamers in Escherichia coli

Autor:	Donald H. Burke, David G. Nickens, James Patterson
Rok vydání:	2003
Předmět:	biology Aptamer Mutant Temperature RNA RNA-dependent RNA polymerase medicine.disease_cause Molecular biology HIV Reverse Transcriptase Reverse transcriptase Report Escherichia coli HIV-1 medicine biology.protein DNA polymerase I Molecular Biology Polymerase
Zdroj:	RNA. 9:1029-1033
ISSN:	1469-9001 1355-8382
DOI:	10.1261/rna.5550103
Popis:	A better understanding of aptamer function in bacteria would help to establish simple model systems for screening RNA–protein interactions within an intracellular context. Escherichia coli DNA polymerase I mutants (Pol Its) fail to grow at 37°C unless an exogenous DNA polymerase such as HIV-1 reverse transcriptase (RT) is expressed within the cell. Here, we show that four RNA aptamers that inhibit HIV-1 RT in vitro block complementation by HIV-1 RT when expressed in vivo. No other essential functions are impaired by aptamer expression at either temperature. Intracellular aptamer RNA concentrations from induced cultures were measured to range from 76 to 180 nM, which is comparable with exogenously expressed HIV-1 RT levels in these cells. RT polymerase activity was reduced to background levels in cell-free extracts prepared from cultures expressing both HIV-1 RT and the 70.28 aptamer, compared with extracts from cultures expressing HIV-1 RT alone. Intracellularly expressed RNA aptamers can thus be used to generate conditional null mutants in bacteria by titrating an essential protein.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3b2109a709a28054f5d6da1a57ddd4d3 https://doi.org/10.1261/rna.5550103 Zobrazit plný text záznamu