Recombinant Coxiella burnetii Uncharacterized protein CBU_1754 (CBU_1754)
Description
Definition and Biological Context
CBU_1754 is a hypothetical protein encoded by C. burnetii, the causative agent of Q fever. It is classified as an LvgA-related protein due to structural similarities with Legionella pneumophila LvgA, a component of the Dot/Icm T4SS apparatus . The recombinant form is expressed in heterologous systems (e.g., E. coli, yeast, insect cells) to study its role in bacterial pathogenesis .
Functional Role in C. burnetii Pathogenesis
CBU_1754 is critical for intracellular replication and maturation of the Coxiella-containing vacuole (CCV):
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Dot/Icm T4SS Interaction: CBU_1754 forms a chaperone complex with C. burnetii proteins IcmS and IcmW, facilitating effector translocation via the T4SS .
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CCV Biogenesis: Deletion of cbu1754 results in smaller CCVs lacking LC3 autophagy markers, impairing bacterial replication .
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Non-Effector Role: Unlike T4SS substrates, CBU_1754 is not translocated into host cells but functions as a structural component of the secretion system .
Recombinant Production and Host Systems
Recombinant CBU_1754 is optimized for yield and functionality across expression systems:
| Host System | Yield | Posttranslational Modifications | Turnaround Time |
|---|---|---|---|
| E. coli | High | Limited | Short |
| Yeast | High | Moderate | Short |
| Insect Cells | Moderate | Extensive (e.g., glycosylation) | Longer |
| Mammalian Cells | Low | Full | Longest |
Translocation Assays
CBU_1754 was not secreted by C. burnetii in CyaA or BlaM reporter assays, contradicting earlier reports of its secretion by L. pneumophila . This highlights species-specific differences in T4SS substrate recognition.
CRISPR Interference (CRISPRi) Data
While CBU0122, CBU1752, CBU1825, and CBU2007 were identified as T4SS substrates promoting CCV biogenesis, CBU_1754 itself was not translocated, reinforcing its role in T4SS machinery .
Implications for Bacterial Pathogenesis
CBU_1754 is indispensable for:
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Effector Translocation: Its interaction with IcmS/IcmW ensures proper T4SS function .
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Host Cell Survival: Mutants exhibit reduced replication, linking CBU_1754 to C. burnetii virulence .
Comparative Analysis of Secretion Studies
Future Directions
Product Specs
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