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A molecular tweezer antagonizes seminal amyloids and HIV infection.

Lump E, Castellano LM, Meier C, Seeliger J, Erwin N, Sperlich B, Stürzel CM, Usmani S, Hammond RM, von Einem J, Gerold G, Kreppel F, Bravo-Rodriguez K, Pietschmann T, Holmes VM, Palesch D, Zirafi O, Weissman D, Sowislok A, Wettig B, Heid C, Kirchhoff F, Weil T, Klärner FG, Schrader T, Bitan G, Sanchez-Garcia E, Winter R, Shorter J, Münch J - Elife (2015)

Bottom Line: In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils.We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism.CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.

ABSTRACT
Semen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides that target viral components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement of viral infection by preventing the formation of virion-amyloid complexes and by directly disrupting the membrane integrity of HIV and other enveloped viruses. We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism. CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself. These combined anti-amyloid and antiviral activities make CLR01 a promising topical microbicide for blocking infection by HIV and other sexually transmitted viruses.

No MeSH data available.


Related in: MedlinePlus

CLR01 does not quench the fluorescence signal of MLV-Gag-YFP particles.MLV-Gag-YFP particles were treated with 300 µM CLR01 or CLR03 for 30 min at 37°C. Thereafter, virions were recovered in the pellet fraction via high-speed centrifugation and aliquots analyzed by confocal microscopy. Z-stack images of the samples revealed a similar amount of fluorescent particles after treatment with CLR01 or CLR03 but altered distribution in the slides.DOI:http://dx.doi.org/10.7554/eLife.05397.015
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fig4s2: CLR01 does not quench the fluorescence signal of MLV-Gag-YFP particles.MLV-Gag-YFP particles were treated with 300 µM CLR01 or CLR03 for 30 min at 37°C. Thereafter, virions were recovered in the pellet fraction via high-speed centrifugation and aliquots analyzed by confocal microscopy. Z-stack images of the samples revealed a similar amount of fluorescent particles after treatment with CLR01 or CLR03 but altered distribution in the slides.DOI:http://dx.doi.org/10.7554/eLife.05397.015

Mentions: Since fibril–virion complexes are already present before ejaculation or form rapidly post emission (Usmani et al., 2014), we next investigated whether CLR01 could displace virions from preformed fibril–virion complexes. Remarkably, CLR01 but not CLR03 substantially reduced the number of virions covering the surface of SEVI, PAP85-120, and SEM1(49-107) fibrils (Figure 4C). We excluded the possibility that CLR01 quenches the fluorescence of YFP-tagged virions by analyzing virions treated with CLR01 or controls by confocal microscopy. We found that treatment of virions with CLR01 did not affect virion fluorescence (Figure 4D) or the number of fluorescent particles (Figure 4—figure supplement 2). However, CLR01-treated virions were dispersed throughout the chamber indicating that their biophysical properties might be altered (Figure 4—figure supplement 2). Our results demonstrate that CLR01 not only prevents the interaction of fibrils with virions but also displaces viral particles from preformed fibril–virion complexes.


A molecular tweezer antagonizes seminal amyloids and HIV infection.

Lump E, Castellano LM, Meier C, Seeliger J, Erwin N, Sperlich B, Stürzel CM, Usmani S, Hammond RM, von Einem J, Gerold G, Kreppel F, Bravo-Rodriguez K, Pietschmann T, Holmes VM, Palesch D, Zirafi O, Weissman D, Sowislok A, Wettig B, Heid C, Kirchhoff F, Weil T, Klärner FG, Schrader T, Bitan G, Sanchez-Garcia E, Winter R, Shorter J, Münch J - Elife (2015)

CLR01 does not quench the fluorescence signal of MLV-Gag-YFP particles.MLV-Gag-YFP particles were treated with 300 µM CLR01 or CLR03 for 30 min at 37°C. Thereafter, virions were recovered in the pellet fraction via high-speed centrifugation and aliquots analyzed by confocal microscopy. Z-stack images of the samples revealed a similar amount of fluorescent particles after treatment with CLR01 or CLR03 but altered distribution in the slides.DOI:http://dx.doi.org/10.7554/eLife.05397.015
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4536748&req=5

fig4s2: CLR01 does not quench the fluorescence signal of MLV-Gag-YFP particles.MLV-Gag-YFP particles were treated with 300 µM CLR01 or CLR03 for 30 min at 37°C. Thereafter, virions were recovered in the pellet fraction via high-speed centrifugation and aliquots analyzed by confocal microscopy. Z-stack images of the samples revealed a similar amount of fluorescent particles after treatment with CLR01 or CLR03 but altered distribution in the slides.DOI:http://dx.doi.org/10.7554/eLife.05397.015
Mentions: Since fibril–virion complexes are already present before ejaculation or form rapidly post emission (Usmani et al., 2014), we next investigated whether CLR01 could displace virions from preformed fibril–virion complexes. Remarkably, CLR01 but not CLR03 substantially reduced the number of virions covering the surface of SEVI, PAP85-120, and SEM1(49-107) fibrils (Figure 4C). We excluded the possibility that CLR01 quenches the fluorescence of YFP-tagged virions by analyzing virions treated with CLR01 or controls by confocal microscopy. We found that treatment of virions with CLR01 did not affect virion fluorescence (Figure 4D) or the number of fluorescent particles (Figure 4—figure supplement 2). However, CLR01-treated virions were dispersed throughout the chamber indicating that their biophysical properties might be altered (Figure 4—figure supplement 2). Our results demonstrate that CLR01 not only prevents the interaction of fibrils with virions but also displaces viral particles from preformed fibril–virion complexes.

Bottom Line: In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils.We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism.CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.

ABSTRACT
Semen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides that target viral components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement of viral infection by preventing the formation of virion-amyloid complexes and by directly disrupting the membrane integrity of HIV and other enveloped viruses. We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism. CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself. These combined anti-amyloid and antiviral activities make CLR01 a promising topical microbicide for blocking infection by HIV and other sexually transmitted viruses.

No MeSH data available.


Related in: MedlinePlus