Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse Transcriptase

The small dumbbell oligonucleotides containing loops of phosphodiester (OL-1), two trimethylene, C3 moieties in each loop (OL-2) and phosphorothioate (OL-3) linkages were synthesized. Incubation of OL-1 and OL-2 with S-1 nuclease generated break down products whereas incubation of OL-3 did not resul...

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Main Author: Ajay Kumar
Format: Article
Language:English
Published: Wiley 2011-01-01
Series:E-Journal of Chemistry
Online Access:http://dx.doi.org/10.1155/2011/584368
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author Ajay Kumar
author_facet Ajay Kumar
author_sort Ajay Kumar
collection DOAJ
description The small dumbbell oligonucleotides containing loops of phosphodiester (OL-1), two trimethylene, C3 moieties in each loop (OL-2) and phosphorothioate (OL-3) linkages were synthesized. Incubation of OL-1 and OL-2 with S-1 nuclease generated break down products whereas incubation of OL-3 did not result in significant cleavage. Their binding to moloney murine leukemia virus reverse transcriptase was evaluated by PAGE band mobility shift assays. The OL-3 bound more strongly to the reverse transcriptase than OL-1 and OL-2. The dissociation constants evaluated using PAGE band mobility shift assays were of the order of 10-7. Investigation of inhibition of RNase H activity of reverse transcriptase showed that the OL-3 is a better inhibitor of the retroviral RNase H activity than both OL-1 and OL-2. Thus OL-3 may be used as RNase H inhibitor. Our studies demonstrated that this particularly designed oligonucleotide (OL-3) displays an IC50 of 25 nM in its inhibition on the reverse transcriptase RNase H activity, a magnitude lower than that of first nucleotide reverse transcriptase of HIV-1, tenofovir, introduced by Gilead Science in the market.
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spelling doaj-art-e5662d1421ad4cb19f82cf22bada179d2025-08-20T02:21:19ZengWileyE-Journal of Chemistry0973-49452090-98102011-01-018262963410.1155/2011/584368Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse TranscriptaseAjay Kumar0Apeejay Stya University, Palwal Sohna Road, Gurgaon, Haryana, IndiaThe small dumbbell oligonucleotides containing loops of phosphodiester (OL-1), two trimethylene, C3 moieties in each loop (OL-2) and phosphorothioate (OL-3) linkages were synthesized. Incubation of OL-1 and OL-2 with S-1 nuclease generated break down products whereas incubation of OL-3 did not result in significant cleavage. Their binding to moloney murine leukemia virus reverse transcriptase was evaluated by PAGE band mobility shift assays. The OL-3 bound more strongly to the reverse transcriptase than OL-1 and OL-2. The dissociation constants evaluated using PAGE band mobility shift assays were of the order of 10-7. Investigation of inhibition of RNase H activity of reverse transcriptase showed that the OL-3 is a better inhibitor of the retroviral RNase H activity than both OL-1 and OL-2. Thus OL-3 may be used as RNase H inhibitor. Our studies demonstrated that this particularly designed oligonucleotide (OL-3) displays an IC50 of 25 nM in its inhibition on the reverse transcriptase RNase H activity, a magnitude lower than that of first nucleotide reverse transcriptase of HIV-1, tenofovir, introduced by Gilead Science in the market.http://dx.doi.org/10.1155/2011/584368
spellingShingle Ajay Kumar
Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse Transcriptase
E-Journal of Chemistry
title Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse Transcriptase
title_full Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse Transcriptase
title_fullStr Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse Transcriptase
title_full_unstemmed Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse Transcriptase
title_short Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse Transcriptase
title_sort small dumbbell oligonucleotides inhibitors of rnase h activity of momulv reverse transcriptase
url http://dx.doi.org/10.1155/2011/584368
work_keys_str_mv AT ajaykumar smalldumbbelloligonucleotidesinhibitorsofrnasehactivityofmomulvreversetranscriptase