Quantitative Infection Dynamics of Cafeteria Roenbergensis Virus

Taylor, Bradford P.; Weitz, Joshua S.; Brussaard, Corina P. D.; Fischer, Matthias

doi:10.3390/v10090468

Cited by 6 publications

(11 citation statements)

References 40 publications

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“…Although CroV encodes some translation components, it lacks ribosomal proteins and therefore depends on host translation. Newly synthesized CroV particles are released as early as 5-6 hours post infection (hpi), while the majority of extracellular virions appear during host cell lysis at 15-22 hpi (Taylor et al , 2018).…”

Section: Introductionmentioning

confidence: 99%

Visualization of giant virus particles using BONCAT labeling and STED microscopy

Berjón-Otero

Duponchel

Hackl

et al. 2020

Preprint

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Giant DNA viruses of the phylum Nucleocytoviricota are being increasingly recognized as important regulators of natural protist populations. However, our knowledge of their infection cycles is still very limited due to a lack of cultured virus-host systems and molecular tools to study them. Here, we apply bioorthogonal noncanonical amino acid tagging (BONCAT) to pulse label the marine heterotrophic flagellate Cafeteria burkhardae during infection with the lytic giant virus CroV. In absence of CroV, we report efficient incorporation of the L-methionine analog L-azidohomoalanine (AHA) into newly synthesized proteins of the methionine prototrophic C. burkhardae. During CroV infection, AHA was predominantly found in viral proteins, and single CroV virions were imaged with stimulated emission depletion (STED) super-resolution microscopy. CroV particles incorporated AHA with 95-100% efficiency while retaining their infectivity, which makes BONCAT/STED a powerful tool to study viral replication cycles in this ecologically relevant marine bacterivore.

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Section: Introductionmentioning

confidence: 99%

Visualization of giant virus particles using BONCAT labeling and STED microscopy

Berjón-Otero

Duponchel

Hackl

et al. 2020

Preprint

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“…Future work at higher MOPs or with different chloroviruses may need to incorporate MOP-dependencies (cf. Brown & Bidle, 2014; Taylor et al ., 2018). Finally, our methods estimated an average burst size, while studies in other viruses have shown that per-cell burst size can vary by orders of magnitude (Kannoly et al ., n.d.; Delbrück, 1945; Timm & Yin, 2012).…”

Section: Discussionmentioning

confidence: 99%

“…By leveraging the distinction between MOP and MOI, the mOSG and mP assays measure more traits than assays based on one or the other (e.g. Van Etten et al ., 1983; Tomaru et al ., 2005; Taylor et al ., 2018). The high-throughput flow cytometry also makes them more efficient than e.g.…”

Section: Discussionmentioning

confidence: 99%

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Life history diversity in a large group of chloroviruses

Lievens

Agarkova

Dunigan

et al. 2022

Preprint

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Microbial viruses affect the ecology and evolution of every environment on earth. Although these effects can play out on a grand scale, to understand them we need to zoom in to virus' life cycle traits. The values of these traits determine a virus' effect on the host population, the trait diversity provides the material for natural selection, and trade-offs between the traits are expected to constrain virus' evolutionary potential. However, due to the difficulty involved in measuring viral life history phenotypes, there are not many studies that characterize their diversity within clades of viruses. In this study, we quantified the life history of 35 chloroviruses from three host range groups (NC64A, Osy, and SAG viruses). We developed methods to efficiently measure adsorption rate, depolarization probability, lysis probability, lysis time, burst size, specific infectivity, and decay. The trait estimates varied between 5 and 75-fold across all the virus strains, with most traits varying substantially both within and across groups. We then used our largest virus group, the NC64A viruses (n = 20), to look for correlations that could signal trade-offs between traits. We found that viruses with high depolarization probabilities also had higher burst sizes, but lower lysis probabilities once depolarized. These correlations were strong, but their mechanistic underpinnings are unclear. We also found a strong correlation between growth rate and survival in the NC64A viruses. Finally, we discovered biphasic or 'tailing' decay in all the chlorovirus groups. The nature and diversity of these traits has large implications for the ecology and evolutionary potential of the chloroviruses, and confirm the importance of studying viral life cycle traits.

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“…In the case of C. burkhardae , the giant virus CroV and the virophage mavirus, the numbers of viral progeny depend on the mode of infection among other factors. CroV-mavirus coinfection (CroV is inhibited by mavirus) and reactivation (both CroV and mavirus replicate, ~130-CroV particles, ~1000 mavirus particles) of the integrated mavirus virophage lead to the replication and release of infectious mavirus particles [ 17 , 18 ]. Coinfection of host cells with CroV and mavirus leads to the inhibition of CroV replication, as mavirus uses the molecular infrastructure of the cytoplasmic CroV factory for its own replication [ 5 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Giant Virus and Virophage Quantification Using Droplet Digital PCR

Arco

Fischer

Becks

2022

Viruses

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Viruses are an abundant component of aquatic systems, but their detection and quantification remain a challenge. Virophages co-replicate with giant viruses in the shared host cell, and can inhibit the production of new giant virus particles, thereby increasing the survival of the infected host population. Here, we present a protocol for Droplet Digital PCR (ddPCR) to quantify simultaneously giant virus and virophage in a mixed sample, enabling the rapid, culture-free and high throughput detection of virus and virophage. As virophage can be present as free virus particles or integrated into the virus host’s genome as well as associated with organic particles, we developed a simple method that enables discrimination between free and particle-associated virophages. The latter include aggregated virophage particles as well as virophage integrated into the host genome. We used, for our experiments, a host-virus-virophage system consisting of Cafeteria burkhardae, CroV and mavirus. Our results show that ddPCR can be an efficient method to quantify virus and virophage, and we discuss potential applications of the method for studying ecological and evolutionary processes of virus and virophages.

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Quantitative Infection Dynamics of Cafeteria Roenbergensis Virus

Cited by 6 publications

References 40 publications

Visualization of giant virus particles using BONCAT labeling and STED microscopy

Visualization of giant virus particles using BONCAT labeling and STED microscopy

Life history diversity in a large group of chloroviruses

Simultaneous Giant Virus and Virophage Quantification Using Droplet Digital PCR

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