Alkoxylalkyl Esters of Nucleotide Analogs Inhibit Polyomavirus DNA Replication and Large T Antigen Activities

Onwubiko, Nichodemus O.; Diaz, Suraya A.; Krečmerová, Marcela; Nasheuer, Heinz-Peter

doi:10.1128/aac.01641-20

Cited by 3 publications

(2 citation statements)

References 81 publications

(141 reference statements)

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“…For EMSA, radioactively labelled ssDNA oligonucleotides (GAPDH ssDNA primer, see above) were incubated with the indicated amounts of proteins in binding buffer (10 mM HEPES-KOH, pH 7.8, 1 mM EDTA and 40 mM NaCl) for 30 min in a volume of 15 lL) and then loaded on a native polyacrylamide gel. The free and protein-bound DNA were separated by EMSAs as previously described [30,38,39]. Autoradiography of the gels was carried out with a Fuji FLA5100 phosphor-imager (Fujifilm, D€ usseldorf, Germany) and free ssDNA and oligomeric Tag-ssDNA complexes were quantified using the program IMAGEQUANT (Fujifilm).…”

Section: Dna Binding Assaysmentioning

confidence: 99%

SV40 T antigen helicase domain regions responsible for oligomerisation regulate Okazaki fragment synthesis initiation

et al. 2022

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The initiation of Okazaki fragment synthesis during cellular DNA replication is a crucial step for lagging strand synthesis, which is carried out by the primase function of DNA polymerase α‐primase (Pol‐prim). Since cellular replication protein A (RPA) prevents primase from starting RNA synthesis on single‐stranded DNA (ssDNA), primase requires auxiliary factors, such as the simian virus 40 (SV40) T antigen (Tag), for the initiation reaction on RPA‐bound ssDNA. Here, we investigated the ability of Tag variants and Tag protein complexes to bind to ssDNA and their resulting effects on the stimulation of Pol‐prim on free and RPA‐bound ssDNA. Atomic force microscopy imaging showed that while Tag131‐627(V350E/P417D) and Tag131‐627(L286D/R567E) (abbreviated as M1 and M2, respectively) could bind to ssDNA as monomers, these monomeric Tags could come together and bind to ssDNA as dimers as well. In a model assay for the initiation of Okazaki fragment synthesis, full‐length Tag SV40 Tag1‐708 and monomeric M2 stimulated DNA synthesis of Pol‐prim on ssDNA and on RPA‐bound ssDNA. In contrast, neither monomeric M1 nor M1‐M2 dimers could stimulate Pol‐prim, on ssDNA or on RPA‐bound ssDNA. Overall, we show that a lack of stimulatory activity of monomeric M1 and M1‐M2 dimers suggests that residues V350 and P417 are not only important for interactions between Tag molecules but also for protein–protein interactions within Okazaki fragment initiation complexes. Thus, we highlight that mutations in M1 are dominant negative with regard to Okazaki fragment initiation.

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Section: Dna Binding Assaysmentioning

confidence: 99%

SV40 T antigen helicase domain regions responsible for oligomerisation regulate Okazaki fragment synthesis initiation

et al. 2022

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“…For example, polyomaviruses use the Tag protein, which binds to viral dsDNA and initiates its replication via cellular polymerases. HDP-CDV was found to be localized to the origin-binding domain of Tag, preventing replication initiation and inhibiting polyomavirus replication [ 78 ].…”

Section: Polymerase Targetingmentioning

confidence: 99%

Recent Advances in Molecular Mechanisms of Nucleoside Antivirals

Kamzeeva,

Aralov,

Alferova

et al. 2023

CIMB

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The search for new drugs has been greatly accelerated by the emergence of new viruses and drug-resistant strains of known pathogens. Nucleoside analogues (NAs) are a prospective class of antivirals due to known safety profiles, which are important for rapid repurposing in the fight against emerging pathogens. Recent improvements in research methods have revealed new unexpected details in the mechanisms of action of NAs that can pave the way for new approaches for the further development of effective drugs. This review accounts advanced techniques in viral polymerase targeting, new viral and host enzyme targeting approaches, and prodrug-based strategies for the development of antiviral NAs.

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Brincidofovir inhibits polyomavirus infection in vivo

Butic,

Katz,

Jin

et al. 2024

mBio

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Polyomaviruses are species-specific DNA viruses that can cause disease in immunocompromised individuals. Despite their role as the causative agents for several diseases, there are no currently approved antivirals for treating polyomavirus infection. Brincidofovir (BCV) is an antiviral approved for the treatment of poxvirus infections and has shown activity against other double-stranded DNA viruses. In this study, we tested the efficacy of BCV against polyomavirus infection in vitro and in vivo using mouse polyomavirus (MuPyV). BCV inhibited virus production in primary mouse kidney cells and brain cortical cells. BCV treatment of cells transfected with MuPyV genomic DNA resulted in a reduction in virus levels, indicating that viral inhibition occurs post-entry. Although in vitro BCV treatment had a limited effect on viral DNA and RNA levels, drug treatment was associated with a reduction in viral protein, raising the possibility that BCV acts post-transcriptionally to inhibit MuPyV infection. In mice, BCV treatment was well tolerated, and prophylactic treatment resulted in a reduction in viral DNA levels and a potent suppression of infectious virus production in the kidney and brain. In mice with chronic polyomavirus infection, therapeutic administration of BCV decreased viremia and reduced infection in the kidney. These data demonstrate that BCV exerts antiviral activity against polyomavirus infection in vivo , supporting further investigation into the use of BCV to treat clinical polyomavirus infections. IMPORTANCE Widespread in the human population and able to persist asymptomatically for the life of an individual, polyomavirus infections cause a significant disease burden in the immunocompromised. Individuals undergoing immune suppression, such as kidney transplant patients or those treated for autoimmune diseases, are particularly at high risk for polyomavirus-associated diseases. Because no antiviral agent exists for treating polyomavirus infections, management of polyomavirus-associated diseases typically involves reducing or discontinuing immunomodulatory therapy. This can be perilous due to the risk of transplant rejection and the potential development of adverse immune reactions. Thus, there is a pressing need for the development of antivirals targeting polyomaviruses. Here, we investigate the effects of brincidofovir, an FDA-approved antiviral, on polyomavirus infection in vivo using mouse polyomavirus. We show that the drug is well-tolerated in mice, reduces infectious viral titers, and limits viral pathology, indicating the potential of brincidofovir as an anti-polyomavirus therapeutic.

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Alkoxylalkyl Esters of Nucleotide Analogs Inhibit Polyomavirus DNA Replication and Large T Antigen Activities

Cited by 3 publications

References 81 publications

SV40 T antigen helicase domain regions responsible for oligomerisation regulate Okazaki fragment synthesis initiation

SV40 T antigen helicase domain regions responsible for oligomerisation regulate Okazaki fragment synthesis initiation

Recent Advances in Molecular Mechanisms of Nucleoside Antivirals

Brincidofovir inhibits polyomavirus infection in vivo

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