Aryl diketoacids (ADK) selectively inhibit duplex DNA-unwinding activity of SARS coronavirus NTPase/helicase

Lee, Chaewoon; J, Lee; Lee, Nara; Jin, Bong-Suk; Jang, Kyoung Jin; Kim, Dong-Eun; Jeong, Young-Keun; Chong, Youhoon

doi:10.1016/j.bmcl.2009.02.010

Cited by 49 publications

(46 citation statements)

References 17 publications

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“…Analysis of amino acid sequence suggests that the SCV helicase has 2 separate domains: (1) a metal-binding domain (MBD) at the N-terminus and (2) a helicase domain (Hel) [22]. A detailed understanding of the biochemical mechanism mediated by SCV helicase became possible [25][26][27][28][29] after its purification [6,30].…”

Section: Characterization Of the Scv Helicase Proteinmentioning

confidence: 99%

“…Further, Lee et al recently identified some aryl diketoacids (ADKs) as chemical inhibitors of SCV helicase. However, in contrast to myricetin and scutellarein, they appeared to inhibit the duplex DNA-unwinding activity but not the ATPase activity of SCV helicase [26]. They have expanded the study by showing that dihydroxychromone derivatives, which are structural analogs of ADKs, can function as chemical inhibitors of the DNA-unwinding activity, but not of the ATPase activity, of SCV helicase [27].…”

Section: Development Of Chemical Inhibitors Of the Scv Helicase Nsp13mentioning

confidence: 99%

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Development of chemical inhibitors of the SARS coronavirus: Viral helicase as a potential target

Keum

Jeong

2012

Biochemical Pharmacology

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Severe acute respiratory syndrome (SARS) was the first pandemic in the 21st century to claim more than 700 lives worldwide. However, effective anti-SARS vaccines or medications are currently unavailable despite being desperately needed to adequately prepare for a possible SARS outbreak. SARS is caused by a novel coronavirus, and one of its components, a viral helicase, is emerging as a promising target for the development of chemical SARS inhibitors. In the following review, we describe the characterization, family classification, and kinetic movement mechanisms of the SARS coronavirus (SCV) helicase-nsP13. We also discuss the recent progress in the identification of novel chemical inhibitors of nsP13 in the context of our recent discovery of the strong inhibition of the SARS helicase by natural flavonoids, myricetin and scutellarein. These compounds will serve as important resources for the future development of anti-SARS medications.

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Section: Characterization Of the Scv Helicase Proteinmentioning

confidence: 99%

Section: Development Of Chemical Inhibitors Of the Scv Helicase Nsp13mentioning

confidence: 99%

Development of chemical inhibitors of the SARS coronavirus: Viral helicase as a potential target

Keum

Jeong

2012

Biochemical Pharmacology

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“…Since the viral helicase has been identified as a potential target for therapy in other viruses due to its indispensability in viral genome replication (10)(11)(12), SCV NTPase/ Helicase (nsP13), which was recently purified and characterized, was suggested as an attractive target for the development of anti-SCV agents (7). Thus, a lot of efforts have been made to identify and test small molecule inhibitors of the SCV helicase as drug candidates (13)(14)(15)(16). RNA and DNA aptamers against SCV helicase were also reported to have an inhibitory effect against nucleic acids unwinding (17,18).…”

Section: Introductionmentioning

confidence: 99%

Cooperative translocation enhances the unwinding of duplex DNA by SARS coronavirus helicase nsP13

Lee

Kwon

Park

et al. 2010

Nucleic Acids Research

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SARS coronavirus encodes non-structural protein 13 (nsP13), a nucleic acid helicase/NTPase belonging to superfamily 1 helicase, which efficiently unwinds both partial-duplex RNA and DNA. In this study, unwinding of DNA substrates that had different duplex lengths and 5′-overhangs was examined under single-turnover reaction conditions in the presence of excess enzyme. The amount of DNA unwound decreased significantly as the length of the duplex increased, indicating a poor in vitro processivity. However, the quantity of duplex DNA unwound increased as the length of the single-stranded 5′-tail increased for the 50-bp duplex. This enhanced processivity was also observed for duplex DNA that had a longer single-stranded gap in between. These results demonstrate that nsP13 requires the presence of a long 5′-overhang to unwind longer DNA duplexes. In addition, enhanced DNA unwinding was observed for gapped DNA substrates that had a 5′-overhang, indicating that the translocated nsP13 molecules pile up and the preceding helicase facilitate DNA unwinding. Together with the propensity of oligomer formation of nsP13 molecules, we propose that the cooperative translocation by the functionally interacting oligomers of the helicase molecules loaded onto the 5′-overhang account for the observed enhanced processivity of DNA unwinding.

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“…Several viral helicases have been used as proven drug targets due to the inhibition of helicase activity in animal models of herpes simplex virus (HSV) and in the treatment of hepatitis C 14,15 . Therefore, much effort has been spent on the development of small-molecule inhibitors and chemicals as drug candidates to inhibit the function of SARS coronavirus helicase nsP13 (SCV nsP13) [16][17][18][19][20] . Helicases are motor proteins that unwind a double-stranded (ds) nucleic acid into two single-stranded (ss) nucleic acids using energy derived from NTP hydrolysis during translocation along a single strand for nucleic acid replication, recombination, and DNA repair [21][22][23][24] .…”

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A high ATP concentration enhances the cooperative translocation of the SARS coronavirus helicase nsP13 in the unwinding of duplex RNA

Jang

Jeong

Kang

et al. 2020

Sci Rep

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Severe acute respiratory syndrome coronavirus nonstructural protein 13 (SCV nsP13), a superfamily 1 helicase, plays a central role in viral RNA replication through the unwinding of duplex RNA and DNA with a 5′ single-stranded tail in a 5′ to 3′ direction. Despite its putative role in viral RNA replication, nsP13 readily unwinds duplex DNA by cooperative translocation. Herein, nsP13 exhibited different characteristics in duplex RNA unwinding than that in duplex DNA. nsP13 showed very poor processivity on duplex RNA compared with that on duplex DNA. More importantly, nsP13 inefficiently unwinds duplex RNA by increasing the 5′-ss tail length. As the concentration of nsP13 increased, the amount of unwound duplex DNA increased and that of unwound duplex RNA decreased. The accumulation of duplex RNA/nsP13 complexes increased as the concentration of nsP13 increased. An increased ATP concentration in the unwinding of duplex RNA relieved the decrease in duplex RNA unwinding. Thus, nsP13 has a strong affinity for duplex RNA as a substrate for the unwinding reaction, which requires increased Atps to processively unwind duplex RnA. our results suggest that duplex RnA is a preferred substrate for the helicase activity of nsP13 than duplex DNA at high ATP concentrations.Severe acute respiratory syndrome (SARS) is an acute respiratory infectious disease caused by a novel coronavirus (SARS-CoV or SCV) that has claimed almost 800 deaths in early 2003 1 . SCV is an enveloped, positive single-stranded RNA virus (or (+) ssRNA virus) with a genome of ~30 kb in length 2,3 . Two-thirds of the SCV genome at the 5′-end comprise replicase genes (orf1ab) encoding 16 nonstructural proteins (nsPs). The replicase genes comprising open reading frames (OFR1a and 1b) are translated into two large replicative polyproteins, pp1ab (~790 kDa) and pp1a (~490 kDa), which are involved with and without ribosomal frameshifting into the −1 frame 4,5 . These two translational polyproteins are processed autoproteolytically by the major viral cysteine proteases M PRO or 3CL PRO to produce 16 non-structural proteins (nsPs), including RNA-dependent RNA polymerases (RdRp, nsP12) and NTPase/helicase (nsP13) 6-8 . These viral replicases are the core of membrane-bound replication-transcription complexes that synthesize the entire viral genome and eight subgenomic mRNAs 9,10 .Because viral helicase is considered to be essential for subsequent viral replication and proliferation, it is an important potential target for antiviral therapy [11][12][13] . In addition, the inhibition of these targets may interfere with the metabolism of the infecting virus without strong side effects in patients. Several viral helicases have been used as proven drug targets due to the inhibition of helicase activity in animal models of herpes simplex virus (HSV) and in the treatment of hepatitis C 14,15 . Therefore, much effort has been spent on the development of small-molecule inhibitors and chemicals as drug candidates to inhibit the function of SARS coronavirus helicase nsP13 (SCV ns...

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Aryl diketoacids (ADK) selectively inhibit duplex DNA-unwinding activity of SARS coronavirus NTPase/helicase

Cited by 49 publications

References 17 publications

Development of chemical inhibitors of the SARS coronavirus: Viral helicase as a potential target

Development of chemical inhibitors of the SARS coronavirus: Viral helicase as a potential target

Cooperative translocation enhances the unwinding of duplex DNA by SARS coronavirus helicase nsP13

A high ATP concentration enhances the cooperative translocation of the SARS coronavirus helicase nsP13 in the unwinding of duplex RNA

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