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Published on: September 28, 2022
Virulent Drexlervirial Bacteriophage MSK, Morphological and Genome Resemblance With Rtp Bacteriophage Inhibits the
Muhammad Saleem Iqbal Khan1, Xiangzheng Gao1, Keying Liang1
1Department of Biochemistry, Cancer Institute of the Second Affiliated Hospital (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education), School of Medicine, Zhejiang University, Hangzhou, China.
Abstract:
Phage-host interactions are likely to have the most critical aspect of phage biology. Phages are the most abundant and ubiquitous infectious acellular entities in the biosphere, where their presence remains elusive. Here, the novel Escherichia coli lytic bacteriophage, named MSK, was isolated from the lysed culture of E. coli C (phix174 host). The genome of phage MSK was sequenced, comprising 45,053 bp with 44.8% G + C composition. In total, 73 open reading frames (ORFs) were predicted, out of which 24 showed a close homology with known functional proteins, including one tRNA-arg; however, the other 49 proteins with no proven function in the genome database were called hypothetical. Electron Microscopy and genome characterization have revealed that MSK phage has a rosette-like tail tip. There were, in total, 46 ORFs which were homologous to the Rtp genome. Among these ORFs, the tail fiber protein with a locus tag of MSK_000019 was homologous to Rtp 43 protein, which determines the host specificity. The other protein, MSK_000046, encodes lipoprotein (cor gene); that protein resembles Rtp 45, responsible for preventing adsorption during cell lysis. Thirteen MSK structural proteins were identified by SDS-PAGE analysis. Out of these, 12 were vital structural proteins, and one was a hypothetical protein. Among these, the protein terminase large (MSK_000072) subunit, which may be involved in DNA packaging and proposed packaging strategy of MSK bacteriophage genome, takes place through headful packaging using the pac-sites. Biosafety assessment of highly stable phage MSK genome analysis has revealed that the phage did not possess virulence genes, which indicates proper phage therapy. MSK phage potentially could be used to inhibit the multidrug-resistant bacteria, including AMP, TCN, and Colistin. Further, a comparative genome and lifestyle study of MSK phage confirmed the highest similarity level (87.18% ANI). These findings suggest it to be a new lytic isolated phage species. Finally, Blast and phylogenetic analysis of the large terminase subunit and tail fiber protein put it in Rtp viruses' genus of family Drexlerviridae.
Insights
A novel lytic bacteriophage, MSK, was isolated from Escherichia coli. Its genome analysis reveals potential for phage therapy against multidrug-resistant bacteria, classifying it as a new species within the Rtp viruses genus.
Area of Science:
- Microbiology
- Virology
- Genomics
Background:
- Bacteriophages are crucial in microbial ecosystems and phage biology.
- Phage-host interactions are central to phage biology.
- Phages are abundant and ubiquitous infectious entities.
Purpose of the Study:
- To isolate and characterize a novel lytic bacteriophage from Escherichia coli.
- To determine the genomic and structural features of the isolated phage.
- To assess the potential of the phage for therapeutic applications.
Main Methods:
- Isolation of bacteriophage MSK from lysed Escherichia coli culture.
- Genome sequencing and bioinformatic analysis (ORF prediction, homology searches).
- Electron microscopy for structural characterization.
- SDS-PAGE for identification of structural proteins.
- Biosafety assessment for virulence genes.
- Comparative genomic analysis (ANI) and phylogenetic analysis.
Main Results:
- Isolation of a novel lytic Escherichia coli bacteriophage, MSK.
- Genome size of 45,053 bp with 44.8% G+C content, predicting 73 ORFs.
- Identification of a rosette-like tail tip structure.
- Homology to Rtp genome, with specific ORFs related to host specificity and cell lysis prevention.
- Identification of 13 structural proteins, including terminase large subunit involved in DNA packaging.
- Absence of virulence genes, indicating safety for phage therapy.
- High genomic similarity (87.18% ANI) to Rtp viruses, suggesting a new species.
- Phylogenetic placement within the Rtp viruses genus, family Drexlerviridae.
Conclusions:
- MSK is a novel lytic bacteriophage species of Escherichia coli.
- The phage possesses structural and genomic features suitable for phage therapy.
- MSK demonstrates potential for inhibiting multidrug-resistant bacteria.
- The findings contribute to understanding phage diversity and therapeutic applications.
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