Structural optimization of a retrograde trafficking inhibitor that protects cells from infections by human polyoma- and papillomaviruses

Carney, Daniel W.; Nelson, Christian D. S.; Ferris, Bennett D.; Stevens, Julia P.; Lipovsky, Alex; Kazakov, Temur; DiMaio, Daniel; Atwood, Walter J.; Sello, Jason K.

doi:10.1016/j.bmc.2014.06.053

Cited by 36 publications

(45 citation statements)

References 24 publications

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“…The parent compound of this derivative was originally identified for its ability to block the entry of bacterial toxins, which require retrograde transport for entry (15). Viruses such as polyomavirus and papillomaviruses require the same retrograde transport for entry into cells, and consequently these compounds effectively blocked the establishment of these viral infections (17,24,25). Efficacy against viruses such as HCMV that do not share this entry pathway has not been previously reported.…”

Section: Discussionmentioning

confidence: 99%

“…Results with our miSTX5 virus indicated that STX5 may be important for properly organizing the cVAC. This would be a unique role for STX5 during virus infections as Retro94 has previously been used to block the entry of several viruses, including polyomavirus and papillomavirus (17), indicating a role for STX5 in the entry of these viruses. Since HCMV does not share the entry pathway of these viruses, we anticipated that Retro94 would not affect HCMV entry.…”

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confidence: 98%

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Potent Inhibition of Human Cytomegalovirus by Modulation of Cellular SNARE Syntaxin 5

Cruz

Streck

Ferguson

et al. 2017

J Virol

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Formation of the cytoplasmic viral assembly compartment (cVAC) is an important step for efficient human cytomegalovirus (HCMV) assembly. To do this, the virus must alter and repurpose the normal cellular balance of membrane and protein flux, a process that is not well understood. Although a recent screen identified three viral proteins essential for cVAC formation, less is known about the contribution of cellular factors. We show that HCMV infection increases the protein level of a cellular trafficking factor, syntaxin 5 (STX5), a member of the syntaxin family of SNARE proteins. STX5 is recruited to the cVAC in infected cells and is required for the efficient production of infectious virions. We find that STX5 is important for normal cVAC morphology and the proper localization of viral proteins. A previously identified inhibitor of trafficking, Retro94, causes the mislocalization of STX5, an altered cVAC morphology, and dispersal of viral proteins. The presence of Retro94 results in severely impaired production of infectious virions, with a decrease as great as 5 logs. We show that this inhibition is conserved among different strains of HCMV and the various cell types that support infection, as well as for murine CMV. Thus, our data identify a key cellular trafficking factor important for supporting HCMV infection.IMPORTANCE Human cytomegalovirus (HCMV) infection causes severe disease and mortality in immunocompromised individuals, including organ transplant and AIDS patients. In addition, infection of a developing fetus may result in lifelong complications such as deafness and learning disabilities. Understanding in detail the processes involved in HCMV replication is important for developing novel treatments. One of these essential processes, assembly of infectious virions, takes places in the cytoplasmic viral assembly compartment. We identify a cellular protein, syntaxin 5, important for generating this compartment, and show that it is required for the efficient production of infectious virions. We also show that a small molecule that disrupts this protein also significantly reduces the amount of infectious virions that are generated. Thus, by pinpointing a cellular protein that is important in the replication cycle of HCMV, we identified a novel target that can be pursued for therapeutic intervention.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 98%

Potent Inhibition of Human Cytomegalovirus by Modulation of Cellular SNARE Syntaxin 5

Cruz

Streck

Ferguson

et al. 2017

J Virol

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“…Recent DHQZ SAR studies have revealed that some SAR analogs can more efficiently block ricin or Shiga toxin transport while others preferentially block transport of papillomavirus and polyomavirus [7][8]. This therefore suggested that there might be subtle but significant differences in the participation of Stx5 in the retrograde transport of various entities.…”

Section: Discussionmentioning

confidence: 99%

Structurally optimized analogs of the retrograde trafficking inhibitor Retro-2cycl limit Leishmania infections

Craig

Huyghues-Despointes

et al. 2017

PLoS Negl Trop Dis

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In infected mammalian cells, Leishmania parasites reside within specialized compartments called parasitophorous vacuoles (LPVs). We have previously shown that Retro-2, a member of a novel class of small retrograde pathway inhibitors caused reduced LPV sizes and lower parasite numbers during experimental L. mexicana sp. infections. The purpose of this study was to determine if structural analogs of Retro-2cycl reported to have superior potency in the inhibition of retrograde pathway-dependent phenomena (i.e., polyomavirus cellular infection by polyomavrius and Shiga toxin trafficking in cells) are also more effective than the parent compound at controlling Leishmania infections. In addition to their effects on LPV development, we show that two optimized analogs of Retro-2cycl, DHQZ 36 and DHQZ 36.1 limit Leishmania amazonensis infection in macrophages at EC50 of 13.63+/-2.58μM and10.57+/-2.66μM, respectively, which is significantly lower than 40.15μM the EC50 of Retro-2cycl. In addition, these analogs caused a reversal in Leishmania induced suppression of IL-6 release by infected cells after LPS activation. Moreover, we show that in contrast to Retro-2cycl that is Leishmania static, the analogs can kill Leishmania parasites in axenic cultures, which is a desirable attribute for any drug to treat Leishmania infections. Together, these studies validate and extend the published structure-activity relationship analyses of Retro-2cycl.

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“…Although Retro-2 has been shown to protect mice against bacterial and plant toxins (27) and infection with enterohemorrhagic Escherichia coli (77), we were unable to demonstrate protective effects of Retro-2 in a mouse model of VACV infection, at least partly due to poor drug solubility. However, Retro-2 derivatives with greatly increased potency are in development (78)(79)(80) and will be worth testing in the future.…”

Section: Discussionmentioning

confidence: 99%

Retrograde Transport from Early Endosomes to the trans -Golgi Network Enables Membrane Wrapping and Egress of Vaccinia Virus Virions

Sivan

Weisberg

Americo

et al. 2016

J Virol

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The anterograde pathway, from the endoplasmic reticulum through the trans-Golgi network to the cell surface, is utilized by trans-membrane and secretory proteins. The retrograde pathway, which directs traffic in the opposite direction, is used following endocytosis of exogenous molecules and recycling of membrane proteins. Microbes exploit both routes: viruses typically use the anterograde pathway for envelope formation prior to exiting the cell, whereas ricin and Shiga-like toxins and some nonenveloped viruses use the retrograde pathway for cell entry. Mining a human genome-wide RNA interference (RNAi) screen revealed a need for multiple retrograde pathway components for cell-to-cell spread of vaccinia virus. We confirmed and extended these results while discovering that retrograde trafficking was required for virus egress rather than entry. Retro-2, a specific retrograde trafficking inhibitor of protein toxins, potently prevented spread of vaccinia virus as well as monkeypox virus, a human pathogen. Electron and confocal microscopy studies revealed that Retro-2 prevented wrapping of virions with an additional double-membrane envelope that enables microtubular transport, exocytosis, and actin polymerization. The viral B5 and F13 protein components of this membrane, which are required for wrapping, normally colocalize in the trans-Golgi network. However, only B5 traffics through the secretory pathway, suggesting that F13 uses another route to the trans-Golgi network. The retrograde route was demonstrated by finding that F13 was largely confined to early endosomes and failed to colocalize with B5 in the presence of Retro-2. Thus, vaccinia virus makes novel use of the retrograde transport system for formation of the viral wrapping membrane. IMPORTANCEEfficient cell-to-cell spread of vaccinia virus and other orthopoxviruses depends on the wrapping of infectious particles with a double membrane that enables microtubular transport, exocytosis, and actin polymerization. Interference with wrapping or subsequent steps results in severe attenuation of the virus. Some previous studies had suggested that the wrapping membrane arises from the trans-Golgi network, whereas others suggested an origin from early endosomes. Some nonenveloped viruses use retrograde trafficking for entry into the cell. In contrast, we provided evidence that retrograde transport from early endosomes to the trans-Golgi network is required for the membrane-wrapping step in morphogenesis of vaccinia virus and egress from the cell. The potent in vitro inhibition of this step by the drug Retro-2 suggests that derivatives with enhanced pharmacological properties might serve as useful antipoxviral agents. V accinia virus (VACV) is the prototype member of the poxvirus family of cytoplasmic double-stranded DNA viruses (1). The first infectious form of VACV, the intracellular mature virion (MV), is a brick-shaped particle consisting of a nucleoprotein core, lateral bodies, and a surrounding lipoprotein membrane (2). A subset of cytoplasmic MVs are wr...

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Structural optimization of a retrograde trafficking inhibitor that protects cells from infections by human polyoma- and papillomaviruses

Cited by 36 publications

References 24 publications

Potent Inhibition of Human Cytomegalovirus by Modulation of Cellular SNARE Syntaxin 5

Potent Inhibition of Human Cytomegalovirus by Modulation of Cellular SNARE Syntaxin 5

Structurally optimized analogs of the retrograde trafficking inhibitor Retro-2cycl limit Leishmania infections

Retrograde Transport from Early Endosomes to the trans -Golgi Network Enables Membrane Wrapping and Egress of Vaccinia Virus Virions

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