Dataset Information

Viral BONCAT: Detection of HPG incorporation into newly synthesized viral proteins via proteomics

ABSTRACT: BONCAT was adapted and tested as a method for directly quantifying viral production in the ocean. To confirm the successful transfer of host-associated HPG-labeled proteins or peptides into marine viruses, we conducted an independent suite of proteomic experiments with cultured systems to directly assess the production of HPG-labeled viral proteins. We used including Emiliania huxleyi strain CCMP374 and its ~200nm coccolithovirus EhV207 as well as E. coli and its ~50 nm virus T7 as virus-host model systems. These specific model systems were chosen because they represent a range of viral particle sizes and their infection dynamics are well characterized. E. huxleyi/EhV207 also represents an ecologically relevant marine virus-host pair.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Emiliania Huxleyi Virus 207 Escherichia Coli Enterobacteria Phage T7 Emiliania Huxleyi

SUBMITTER: Michael Sweredoski

LAB HEAD: Victoria Orphan

PROVIDER: PXD006580 | Pride | 2017-11-27

REPOSITORIES: Pride

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Publications

Interrogating marine virus-host interactions and elemental transfer with BONCAT and nanoSIMS-based methods.

Pasulka Alexis L AL Thamatrakoln Kimberlee K Kopf Sebastian H SH Guan Yunbin Y Poulos Bonnie B Moradian Annie A Sweredoski Michael J MJ Hess Sonja S Sullivan Mathew B MB Bidle Kay D KD Orphan Victoria J VJ

Environmental microbiology 20171214 2

While the collective impact of marine viruses has become more apparent over the last decade, a deeper understanding of virus-host dynamics and the role of viruses in nutrient cycling would benefit from direct observations at the single-virus level. We describe two new complementary approaches - stable isotope probing coupled with nanoscale secondary ion mass spectrometry (nanoSIMS) and fluorescence-based biorthogonal non-canonical amino acid tagging (BONCAT) - for studying the activity and bioge ...[more]

PMID: 29159966

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Dataset Information

Viral BONCAT: Detection of HPG incorporation into newly synthesized viral proteins via proteomics

Publications

Interrogating marine virus-host interactions and elemental transfer with BONCAT and nanoSIMS-based methods.

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