Journal
CHEMBIOCHEM
Volume 9, Issue 18, Pages 3037-3045Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.200800491
Keywords
antiviral agents; aptamers; DNA structures; g-quadruplexes; nuclease resistance
Funding
- Hong Kong Research Grants Council (RGC) [HKU 7589105M]
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The helicase from severe acute respiratory syndrome coronavirus (SARS-COV) possesses NTPase, duplex RNA/DNA-unwinding and RNA-capping activities that are essential for viral replication and proliferation. Here, we have isolated DNA aptamers against the SARS-CoV helicase from a combinatorial DNA library. These aptamers show two distinct classes of secondary structure, G-quadruplex and non-G-quadruplex, as shown by circular dichroism and gel electrophoresis. All of the aptamers that were selected stimulated ATPase activity of the SARS-CoV helicase with low-nanomolar apparent K-m values. Intriguingly, only the non-G-quadruplex aptamers showed specific inhibition of helicase activities, whereas the G-quadruplex aptamers did not inhibit helicase activities. The non-G-quadruplex aptamer with the strongest inhibitory potency was modified at the 3'-end with biotin or Inverted thymidine, and the modification increased its stability in serum, particularly for the inverted thymidine modification. Structural diversity in selection coupled to post-selection stabilisation has provided new insights into the aptamers that were selected for a helicase target. These aptamers ore being further developed to inhibit SARS-CoV replication.
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