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Updated: Dec 27, 2025

Novel RNA-Binding Proteins Isolation by the RaPID Methodology
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The endoplasmic reticulum-associated mRNA-binding proteins ERBP1 and ERBP2 interact in bloodstream-form Trypanosoma

Kathrin Bajak1,2, Kevin Leiss1, Christine E Clayton1

  • 1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Heidelberg, Germany.

Peerj
|February 26, 2020
PubMed

Insights

Trypanosoma brucei ERBP1, an endoplasmic reticulum-associated protein, binds to ribosomal protein mRNAs. Loss of ERBP1 initially impacts growth, but the parasite adapts, suggesting complex post-transcriptional regulation in kinetoplastids.

Area of Science:

  • Molecular Biology
  • Parasitology
  • Gene Expression Regulation

Background:

  • Kinetoplastids utilize post-transcriptional mechanisms for gene expression control.
  • RNA-binding proteins are crucial for mRNA processing, including splicing, translation, and decay.
  • Trypanosoma brucei ERBP1 and ERBP2 are identified mRNA-binding proteins lacking canonical RNA-binding domains.

Purpose of the Study:

  • To investigate the cellular localization and function of Trypanosoma brucei ERBP1.
  • To determine the interaction between ERBP1 and ERBP2.
  • To identify the specific mRNAs bound by ERBP1.

Main Methods:

  • Immunofluorescence to determine ERBP1 localization.
  • Co-immunoprecipitation to assess ERBP1-ERBP2 interaction.
  • Affinity pull-down assays with tagged ERBP1 to identify bound mRNAs.
  • Growth curve analysis of T. brucei lacking ERBP1.

Main Results:

  • ERBP1 localizes to the endoplasmic reticulum and interacts with ERBP2 in vivo.
  • Loss of ERBP1 causes an initial growth defect in bloodstream-form T. brucei, with recovery upon prolonged cultivation.
  • ERBP1 preferentially binds to ribosomal protein mRNAs.
  • ERBP1 shares similarities with Saccharomyces cerevisiae Bfr1 but does not bind secreted protein mRNAs or recruit to stress granules.

Conclusions:

  • ERBP1 is an endoplasmic reticulum-associated protein involved in post-transcriptional regulation of gene expression in T. brucei.
  • ERBP1's preferential binding to ribosomal protein mRNAs suggests a role in regulating protein synthesis.
  • The adaptive growth recovery indicates compensatory mechanisms in T. brucei gene regulation.
Keywords:
Bfr1Post-transcriptional regulationRNA-binding proteinsRibosomal mRNATrypanosoma brucei

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