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Supersize me: Cronobacter sakazakii phage GAP32
Reza Abbasifar1, Mansel W Griffiths1, Parviz M Sabour2
1Canadian Research Institute for Food Safety, University of Guelph, Guelph, ON, Canada N1G 2W1.
Abstract:
Cronobacter sakazakii is a Gram-negative pathogen found in milk-based formulae that causes infant meningitis. Bacteriophages have been proposed to control bacterial pathogens; however, comprehensive knowledge about a phage is required to ensure its safety before clinical application. We have characterized C. sakazakii phage vB_CsaM_GAP32 (GAP32), which possesses the second largest sequenced phage genome (358,663bp). A total of 571 genes including 545 protein coding sequences and 26 tRNAs were identified, thus more genes than in the smallest bacterium, Mycoplasma genitalium G37. BLASTP and HHpred searches, together with proteomic analyses reveal that only 23.9% of the putative proteins have defined functions. Some of the unique features of this phage include: a chromosome condensation protein, two copies of the large subunit terminase, a predicted signal-arrest-release lysin; and an RpoD-like protein, which is possibly involved in the switch from immediate early to delayed early transcription. Its closest relatives are all extremely large myoviruses, namely coliphage PBECO4 and Klebsiella phage vB_KleM-RaK2, with whom it shares approximately 44% homologous proteins. Since the homologs are not evenly distributed, we propose that these three phages belong to a new subfamily.
Insights
We characterized Cronobacter sakazakii phage GAP32, revealing its massive genome and unique genes. This research is crucial for understanding phage safety and potential applications in controlling bacterial pathogens.
Area of Science:
- Microbiology
- Virology
- Genomics
Background:
- Cronobacter sakazakii is a pathogen in infant formula, causing meningitis.
- Bacteriophages are potential antimicrobial agents, but safety knowledge is vital.
- Phage characterization is essential for clinical safety assessment.
Purpose of the Study:
- To characterize the Cronobacter sakazakii phage vB_CsaM_GAP32 (GAP32).
- To assess the genomic features and potential functions of GAP32.
- To understand the evolutionary relationships of GAP32 with other phages.
Main Methods:
- Genome sequencing and analysis of GAP32.
- Bioinformatic searches (BLASTP, HHpred) for protein homology.
- Proteomic analysis to identify protein functions.
Main Results:
- GAP32 has the second-largest sequenced phage genome (358,663bp) with 571 genes.
- Only 23.9% of GAP32's proteins have known functions, indicating novel biology.
- Unique features include a chromosome condensation protein and a signal-arrest-release lysin.
- GAP32 shares ~44% homologous proteins with coliphage PBECO4 and Klebsiella phage vB_KleM-RaK2.
Conclusions:
- GAP32 represents a novel, large myovirus with unique genetic elements.
- The genomic data suggests GAP32 and its relatives may form a new phage subfamily.
- Further research is needed to elucidate the functions of unknown GAP32 proteins for safety assessment.
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