Cited by 138Trypanosoma species research and implicationsBiotin and Related StudiesGenetic and Kidney Cyst Diseases

Novel Bilobe Components in Trypanosoma brucei Identified Using Proximity-Dependent Biotinylation

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Morriswood, Brooke, Havlicek, Katharina, Demmel, Lars, Yavuz, Sevil, Sealey-Cardona, Marco, Vidilaseris, Keni, Anrather, Dorothea, Kostan, Julius +3 more·2013·Eukaryotic Cell

Abstract

The trypanosomes are a family of parasitic protists of which the African trypanosome, Trypanosoma brucei , is the best characterized. The complex and highly ordered cytoskeleton of T. brucei has been shown to play vital roles in its biology but remains difficult to study, in large part owing to the intractability of its constituent proteins. Existing methods of protein identification, such as bioinformatic analysis, generation of monoclonal antibody panels, proteomics, affinity purification, and yeast two-hybrid screens, all have drawbacks. Such deficiencies—troublesome proteins and technical limitations—are common not only to T. brucei but also to many other protists, many of which are even less well studied. Proximity-dependent biotin identification (BioID) is a recently developed technique that allows forward screens for interaction partners and near neighbors in a native environment with no requirement for solubility in nonionic detergent. As such, it is extremely well suited to the exploration of the cytoskeleton. In this project, BioID was adapted for use in T. brucei . The trypanosome bilobe, a discrete cytoskeletal structure with few known protein components, represented an excellent test subject. Use of the bilobe protein TbMORN1 as a probe resulted in the identification of seven new bilobe constituents and two new flagellum attachment zone proteins. This constitutes the first usage of BioID on a largely uncharacterized structure, and demonstrates its utility in identifying new components of such a structure. This remarkable success validates BioID as a new tool for the study of unicellular eukaryotes in particular and the eukaryotic cytoskeleton in general.

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  • BioID has been used extensively in vitro to define interacting protein partners and proteins in subcellular compartments, but its application in vivo is limited.

    “BioID has been used extensively in vitro [162–166] to define interacting protein partners and proteins in subcellular compartments, but its application in vivo is limited.”

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Authors
Morriswood, B., Havlicek, K., Demmel, L., Yavuz, S., Sealey-Cardona, M., Vidilaseris, K., Anrather, D., Kostan, J., Djinović-Carugo, K., Roux, K. J., & Warren, G.
Affiliations
  • University of Vienna (AT)
  • Max F. Perutz Laboratories, University of Vienna and Medical University of Vienna, Vienna, Austria
  • Department for Structural and Computational Biology, Max F. Perutz Laboratories, University of Vienna, Vienna, Austria
  • Sanford Research (US)
Venue
Eukaryotic Cell
Published
2013-02
Volume
12
Issue
2
Pages
356-367
ISSN
  • 1535-9778 (ISSN-L)
  • 1535-9786
OpenAlex
W2168967957
Cited by
138
References
30
Citations by year
2026: 7 · 2025: 4 · 2024: 3 · 2023: 6 · 2022: 7 · 2021: 11 · 2020: 13 · 2019: 14 · 2018: 11 · 2017: 12 · 2016: 15 · 2015: 15 · 2014: 11 · 2013: 8 · 2012: 1
FWCI
8.35
Type
journal article
Language
en
Open access
Locations
  • Eukaryotic Cell (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Health Sciences › Medicine › Epidemiology › Trypanosoma species research and implications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic and Kidney Cyst Diseases
Keywords
Trypanosoma brucei, cytoskeleton, proximity biotinylation, BioID, flagellum attachment zone, protein-protein interaction, protein identification, trypanosomes
MeSH terms
Cytoskeletal Proteins, Protein Binding, Trypanosoma brucei brucei, Protozoan Proteins, Biotinylation, Protein Transport, Protein Interaction Mapping

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