Development of TEM-1 β-lactamase based protein translocation assay for identification of Anaplasma phagocytophilum

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Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions
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Analysis of Yersinia enterocolitica Effector Translocation into Host Cells Using Beta-lactamase Effector Fusions

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Jiafeng Zhu1, Meiling He1, Wenting Xu1

  • 1Department of Microbiology, College of Medicine, Soochow University, Suzhou, Jiangsu Province, 215123, China.

Scientific Reports
|March 14, 2019
PubMed

Insights

This study introduces a new assay to identify bacterial proteins that Anaplasma phagocytophilum uses to infect host cells. This helps understand human granulocytic anaplasmosis (HGA) pathogenesis.

Area of Science:

  • Microbiology
  • Cell Biology
  • Pathogenesis

Background:

  • Anaplasma phagocytophilum causes human granulocytic anaplasmosis (HGA).
  • This bacterium is an obligate intracellular pathogen with a small genome, requiring host cell manipulation for replication.
  • The type IV secretion system (T4SS) is crucial for translocating bacterial effectors into host cells.

Purpose of the Study:

  • To develop a novel protein translocation assay for identifying Anaplasma phagocytophilum T4SS effectors.
  • To characterize the function and localization of identified T4SS effectors.

Main Methods:

  • Development of a TEM-1 β-lactamase based protein translocation assay.
  • Application of the assay to screen for A. phagocytophilum T4SS effectors.
  • Localization studies of identified effectors in transfected cells.

Main Results:

  • A novel protein translocation assay was successfully developed.
  • The hypothetical protein APH0215 was identified as an Anaplasma phagocytophilum T4SS effector.
  • APH0215 was found to interact with the trans-Golgi network in host cells.

Conclusions:

  • The developed TEM-1 β-lactamase assay is effective for identifying T4SS effectors of Anaplasma phagocytophilum.
  • APH0215 is a novel T4SS effector that interacts with the trans-Golgi network.
  • This research facilitates the identification of T4SS effectors and enhances understanding of HGA pathogenesis.

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