Bacteriophage Bioinformatics

A special issue of Viruses (ISSN 1999-4915). This special issue belongs to the section "Bacterial Viruses".

Deadline for manuscript submissions: closed (30 September 2022) | Viewed by 19946

Special Issue Editors


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Guest Editor
Department of Biological Sciences, Birkbeck University of London, London, UK
Interests: cryo EM; bioinformatics; structural methods; viruses; bacteriophages; portal motors; structure/function relationship
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Guest Editor
Department of Molecular and Cellular Biochemistry, Director of the Electron Microscopy Center, Indiana University, Bloomington, IN, USA
Interests: bacteriophage; virus; virus assembly; DNA packaging; molecular motors; ATPase; ASCE; structure; cryo-electron microscopy; biophysics
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The continued rise of antibiotic resistance to critical heights together with an increasing number of bacterial pathogens harkens back to the pre-penicillin era when producing new antibiotics was an urgent societal need. While promising antimicrobial agents were identified nearly a century ago and long before the discovery of penicillin, their therapeutic potential was never realized and they were rarely used clinically. Indeed, the first such agents were identified as bacteriophages.  However, at that time researchers had neither the proper equipment nor a sufficient understanding of bacteriophage biology to realize the antibiotic potential of phages, possibilities to do high throughput screening (HTS) of protein/ligand interactions, quantitative methods of comparative analysis, which will provide information about these agents, their activity and specificity.  Presently, the situation has radically changed; complementary structural, biochemical, and computational efforts have provided significant insight into how bacteriophages recognize and destroy their bacterial targets.  Indeed, hybrid approaches combining structural methods, bioinformatics analysis of sequence databases, biophysical characterization of phage interactions with various  ligands/proteins and computational statistics with high throughput biochemical methods have provided a wealth of information regarding phage-pathogen interactions, the functions of phage-encoded proteins, and the dependence of phage populations on animal or human microbiomes. As a result, phage therapy is beginning to become a more broadly used treatment against clinically challenging bacterial infections. However, despite these successful efforts, improvements in the analyses and comparison of various components of bacteriophages and their interactions with pathogenic hosts are necessary to understand the principles of regulation of bacteriophage activity. In this context, hybrid approaches that include bioinformatics, biophysical, biochemical, single molecule, and microbiological approaches have the potential to accelerate the development of phage-based therapeutics and usher in a new era antibiotics.

This Special Issue of Viruses will highlight innovations in the application of hybrid methods to analyze bacteriophage activity and function and in the search for bacteriophages that can be rationally engineered to function as powerful antibiotic therapeutics. Approaches that can be used to identify phages with therapeutic potential and/or to create  synthetic phages with desired antibiotic properties have the potential to transform the emerging field of phage therapy. We encourage submissions of reviews or original research manuscripts addressing the following topics:

1)   Tools for the analysis of phage genomes and/or comparative assessments of available software packages.
2)   How prediction bacteriophage type (virulent vs. temperate) can be accomplished via computational analysis of phage genomes. Classification of phages based on their genome sequences
3)   Applications of bioinformatics in verification and classification of the phage proteins
4)   Optimization of high throughput approaches in the analysis of phages utilizing bioinformatics approaches.
5)   Methods to analyze phage-bacterium interactions analysis and links to their life state: virulent or temperate.
6)    Design, and applications of synthetic phages in pharmaceutics and food industries.

Prof. Dr. Elena Orlova
Dr. Marc C. Morais
Guest Editors

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Keywords

  • viruses
  • bacteriophage
  • bacteriophage classification
  • genome
  • bioinformatics
  • computational tools
  • structural organisation
  • infectivity
  • function
  • structural methods
  • electron microscopy
  • X-ray
  • NMR
  • high throughput screening

Related Special Issue

Published Papers (9 papers)

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Research

19 pages, 6114 KiB  
Article
Structure and Function of Hoc—A Novel Environment Sensing Device Encoded by T4 and Other Bacteriophages
by Andrei Fokine, Mohammad Zahidul Islam, Qianglin Fang, Zhenguo Chen, Lei Sun and Venigalla B. Rao
Viruses 2023, 15(7), 1517; https://doi.org/10.3390/v15071517 - 7 Jul 2023
Cited by 3 | Viewed by 2156
Abstract
Bacteriophage T4 is decorated with 155 180 Å-long fibers of the highly antigenic outer capsid protein (Hoc). In this study, we describe a near-atomic structural model of Hoc by combining cryo-electron microscopy and AlphaFold structure predictions. It consists of a conserved C-terminal capsid-binding [...] Read more.
Bacteriophage T4 is decorated with 155 180 Å-long fibers of the highly antigenic outer capsid protein (Hoc). In this study, we describe a near-atomic structural model of Hoc by combining cryo-electron microscopy and AlphaFold structure predictions. It consists of a conserved C-terminal capsid-binding domain attached to a string of three variable immunoglobulin (Ig)-like domains, an architecture well-preserved in hundreds of Hoc molecules found in phage genomes. Each T4-Hoc fiber attaches randomly to the center of gp23* hexameric capsomers in one of the six possible orientations, though at the vertex-proximal hexamers that deviate from 6-fold symmetry, Hoc binds in two preferred orientations related by 180° rotation. Remarkably, each Hoc fiber binds to all six subunits of the capsomer, though the interactions are greatest with three of the subunits, resulting in the off-centered attachment of the C-domain. Biochemical analyses suggest that the acidic Hoc fiber (pI, ~4–5) allows for the clustering of virions in acidic pH and dispersion in neutral/alkaline pH. Hoc appears to have evolved as a sensing device that allows the phage to navigate its movements through reversible clustering–dispersion transitions so that it reaches its destination, the host bacterium, and persists in various ecological niches such as the human/mammalian gut. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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12 pages, 1278 KiB  
Article
Phage-Host Prediction Using a Computational Tool Coupled with 16S rRNA Gene Amplicon Sequencing
by Harilanto Felana Andrianjakarivony, Yvan Bettarel, Fabrice Armougom and Christelle Desnues
Viruses 2023, 15(1), 76; https://doi.org/10.3390/v15010076 - 27 Dec 2022
Viewed by 2208
Abstract
Metagenomics studies have revealed tremendous viral diversity in aquatic environments. Yet, while the genomic data they have provided is extensive, it is unannotated. For example, most phage sequences lack accurate information about their bacterial host, which prevents reliable phage identification and the investigation [...] Read more.
Metagenomics studies have revealed tremendous viral diversity in aquatic environments. Yet, while the genomic data they have provided is extensive, it is unannotated. For example, most phage sequences lack accurate information about their bacterial host, which prevents reliable phage identification and the investigation of phage–host interactions. This study aimed to take this knowledge further, using a viral metagenomic framework to decipher the composition and diversity of phage communities and to predict their bacterial hosts. To this end, we used water and sediment samples collected from seven sites with varying contamination levels in the Ebrié Lagoon in Abidjan, Ivory Coast. The bacterial communities were characterized using the 16S rRNA metabarcoding approach, and a framework was developed to investigate the virome datasets that: (1) identified phage contigs with VirSorter and VIBRANT; (2) classified these contigs with MetaPhinder using the phage database (taxonomic annotation); and (3) predicted the phages’ bacterial hosts with a machine learning-based tool: the Prokaryotic Virus-Host Predictor. The findings showed that the taxonomic profiles of phages and bacteria were specific to sediment or water samples. Phage sequences assigned to the Microviridae family were widespread in sediment samples, whereas phage sequences assigned to the Siphoviridae, Myoviridae and Podoviridae families were predominant in water samples. In terms of bacterial communities, the phyla Latescibacteria, Zixibacteria, Bacteroidetes, Acidobacteria, Calditrichaeota, Gemmatimonadetes, Cyanobacteria and Patescibacteria were most widespread in sediment samples, while the phyla Epsilonbacteraeota, Tenericutes, Margulisbacteria, Proteobacteria, Actinobacteria, Planctomycetes and Marinimicrobia were most prevalent in water samples. Significantly, the relative abundance of bacterial communities (at major phylum level) estimated by 16S rRNA metabarcoding and phage-host prediction were significantly similar. These results demonstrate the reliability of this novel approach for predicting the bacterial hosts of phages from shotgun metagenomic sequencing data. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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11 pages, 1944 KiB  
Article
Two Novel Yersinia pestis Bacteriophages with a Broad Host Range: Potential as Biocontrol Agents in Plague Natural Foci
by Haixiao Jin, Youhong Zhong, Yiting Wang, Chuanyu Zhang, Jin Guo, Xiaona Shen, Cunxiang Li, Ying Huang, Haoming Xiong, Peng Wang and Wei Li
Viruses 2022, 14(12), 2740; https://doi.org/10.3390/v14122740 - 8 Dec 2022
Cited by 1 | Viewed by 1478
Abstract
Bacteriophages (phages) have been successfully used as disinfectors to kill bacteria in food and the environment and have been used medically for curing human diseases. The objective of this research was to elucidate the morphological and genomic characteristics of two novel Yersinia pestis [...] Read more.
Bacteriophages (phages) have been successfully used as disinfectors to kill bacteria in food and the environment and have been used medically for curing human diseases. The objective of this research was to elucidate the morphological and genomic characteristics of two novel Yersinia pestis phages, vB_YpeM_ MHS112 (MHS112) and vB_YpeM_GMS130 (GMS130), belonging to the genus Gaprivervirus, subfamily Tevenvirinae, family Myoviridae. Genome sequencing showed that the sizes of MHS112 and GMS130 were 170507 and 168552 bp, respectively. A total of 303 and 292 open reading frames with 2 tRNA and 3 tRNA were predicted in MHS112 and GMS130, respectively. The phylogenetic relationships were analysed among the two novel Y. pestis phages, phages in the genus Gaprivervirus, and several T4-like phages infecting the Yersinia genus. The bacteriophage MHS112 and GMS130 exhibited a wider lytic host spectrum and exhibited comparative temperature and pH stability. Such features signify that these phages do not need to rely on Y. pestis as their host bacteria in the ecological environment, while they could be based on more massive Enterobacteriales species to propagate and form ecological barriers against Y. pestis pathogens colonised in plague foci. Such characteristics indicated that the two phages have potential as biocontrol agents for eliminating the endemics of animal plague in natural plague foci. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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15 pages, 2254 KiB  
Article
Isolation, Characterization, and Genome Analysis of a Novel Bacteriophage, Escherichia Phage vB_EcoM-4HA13, Representing a New Phage Genus in the Novel Phage Family Chaseviridae
by Janet T. Lin, Sarah Kirst, Stevan Cucić, Alexandra Klem, Yi-Min She, Andrew M. Kropinski and Hany Anany
Viruses 2022, 14(11), 2356; https://doi.org/10.3390/v14112356 - 26 Oct 2022
Cited by 2 | Viewed by 2271
Abstract
Shiga toxin-producing Escherichia coli (STEC) is one of the leading causes of foodborne illnesses in North America and can lead to severe symptoms, with increased fatality risk for young children. While E. coli O157:H7 remains the dominant STEC serotype associated with foodborne outbreaks, [...] Read more.
Shiga toxin-producing Escherichia coli (STEC) is one of the leading causes of foodborne illnesses in North America and can lead to severe symptoms, with increased fatality risk for young children. While E. coli O157:H7 remains the dominant STEC serotype associated with foodborne outbreaks, there has been an increasing number of non-O157 STEC outbreaks in recent years. For the food industry, lytic bacteriophages offer an organic, self-limiting alternative to pathogen reduction—one that could replace or reduce the use of chemical and physical food processing methods. From EHEC-enriched sewage, we isolated a novel bacteriophage, vB_EcoM-4HA13 (4HA13). Phenotypic characterizations revealed 4HA13 to possess a myoviral morphotype, with a high specificity to non-motile O111 serotype, and a long latent period (90 min). Through genomic analyses, this 52,401-bp dsDNA phage was found to contain 81 CDS, but no detectable presence of antibiotic resistance, integrase, or virulence genes. A BLASTn search for each of the identified 81 CDS yielded homologues with low levels of similarity. Comparison of RNA polymerase and terminase large subunit amino acid sequences led to the proposal and acceptance of a new bacteriophage family, Chaseviridae, with 4HA13 representing a new species and genus. The discovery of this phage has broadened our current knowledge of bacteriophage diversity. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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15 pages, 7768 KiB  
Article
Forty Years without Family: Three Novel Bacteriophages with High Similarity to SPP1 Reveal Decades of Evolutionary Stasis since the Isolation of Their Famous Relative
by Véronique A. Delesalle, Brianne E. Tomko, Albert C. Vill, Katherine B. Lichty and Greg P. Krukonis
Viruses 2022, 14(10), 2106; https://doi.org/10.3390/v14102106 - 23 Sep 2022
Cited by 2 | Viewed by 1500
Abstract
SPP1, an extensively studied bacteriophage of the Gram-positive Bacillus subtilis, is a model system for the study of phage–host interactions. Despite progress in the isolation and characterization of Bacillus phages, no previously fully sequenced phages have shared more than passing genetic similarity [...] Read more.
SPP1, an extensively studied bacteriophage of the Gram-positive Bacillus subtilis, is a model system for the study of phage–host interactions. Despite progress in the isolation and characterization of Bacillus phages, no previously fully sequenced phages have shared more than passing genetic similarity to SPP1. Here, we describe three virulent phages very similar to SPP1; SPP1 has greater than 80% nucleotide sequence identity and shares more that 85% of its protein coding genes with these phages. This is remarkable, given more than 40 years between the isolation of SPP1 and these phages. All three phages have somewhat larger genomes and more genes than SPP1. We identified a new putative gene in SPP1 based on a conserved sequence found in all phages. Gene conservation connotes purifying selection and is observed in structural genes and genes involved in DNA metabolism, but also in genes of unknown function, suggesting an important role in phage survival independent of the environment. Patterns of divergence point to genes or gene domains likely involved in adaptation to diverse hosts or different environments. Ultimately, comparative genomics of related phages provides insight into the long-term selective pressures that affect phage–bacteria interactions and alter phage genome content. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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7 pages, 481 KiB  
Communication
Comparative Genomics of Closely-Related Gordonia Cluster DR Bacteriophages
by Cyril J. Versoza, Abigail A. Howell, Tanya Aftab, Madison Blanco, Akarshi Brar, Elaine Chaffee, Nicholas Howell, Willow Leach, Jackelyn Lobatos, Michael Luca, Meghna Maddineni, Ruchira Mirji, Corinne Mitra, Maria Strasser, Saige Munig, Zeel Patel, Minerva So, Makena Sy, Sarah Weiss and Susanne P. Pfeifer
Viruses 2022, 14(8), 1647; https://doi.org/10.3390/v14081647 - 27 Jul 2022
Viewed by 2120
Abstract
Bacteriophages infecting bacteria of the genus Gordonia have increasingly gained interest in the scientific community for their diverse applications in agriculture, biotechnology, and medicine, ranging from biocontrol agents in wastewater management to the treatment of opportunistic pathogens in pulmonary disease patients. However, due [...] Read more.
Bacteriophages infecting bacteria of the genus Gordonia have increasingly gained interest in the scientific community for their diverse applications in agriculture, biotechnology, and medicine, ranging from biocontrol agents in wastewater management to the treatment of opportunistic pathogens in pulmonary disease patients. However, due to the time and costs associated with experimental isolation and cultivation, host ranges for many bacteriophages remain poorly characterized, hindering a more efficient usage of bacteriophages in these areas. Here, we perform a series of computational genomic inferences to predict the putative host ranges of all Gordonia cluster DR bacteriophages known to date. Our analyses suggest that BiggityBass (as well as several of its close relatives) is likely able to infect host bacteria from a wide range of genera—from Gordonia to Nocardia to Rhodococcus, making it a suitable candidate for future phage therapy and wastewater treatment strategies. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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13 pages, 4451 KiB  
Article
Characterization and Genomic Analysis of Novel Vibrio parahaemolyticus Phage vB_VpaP_DE10
by Yuanming Ye, Hanfang Chen, Qiaolan Huang, Shixuan Huang, Jiaxin He, Jumei Zhang, Qingping Wu, Xueling Li, Wenfeng Hu and Meiyan Yang
Viruses 2022, 14(8), 1609; https://doi.org/10.3390/v14081609 - 23 Jul 2022
Cited by 9 | Viewed by 2208
Abstract
In the present study, a novel lytic Vibrio parahaemolyticus phage, vB_VpaP_DE10, was isolated from sewage samples collected in Guangzhou city, China. Transmission electron microscopy revealed that phage vB_VpaP_DE10 has an icosahedral head (52.4 ± 2.5 nm) and a short non-contracted tail (21.9 ± [...] Read more.
In the present study, a novel lytic Vibrio parahaemolyticus phage, vB_VpaP_DE10, was isolated from sewage samples collected in Guangzhou city, China. Transmission electron microscopy revealed that phage vB_VpaP_DE10 has an icosahedral head (52.4 ± 2.5 nm) and a short non-contracted tail (21.9 ± 1.0 nm). Phage vB_VpaP_DE10 lysed approximately 31% (8/26) of the antibiotic-resistant V. parahaemolyticus strains tested. A one-step growth curve showed that phage vB_VpaP_DE10 has a relatively long latency time of 25 min and a burst size of ~19 PFU per cell. The genome of phage vB_VpaP_DE10 is a 42,871-bp-long dsDNA molecule with a G + C content of 49.19% and is predicted to contain 46 open reading frames, 26 of which are predicted to be related to functions such as phage structure, packaging, host lysis, and DNA metabolism. Sequence comparisons suggested that vB_VpaP_DE10 is a member of the genus Maculvirus within the family Autographiviridae. Morphological and genomic analysis indicated that vB_VpaP_DE10 is a novel V. parahaemolyticus phage. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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15 pages, 3431 KiB  
Article
Isolation and Characterization of a Lytic Vibrio parahaemolyticus Phage vB_VpaP_GHSM17 from Sewage Samples
by Xunru Liang, Yuhang Wang, Bin Hong, Yanmei Li, Yi Ma and Jufang Wang
Viruses 2022, 14(8), 1601; https://doi.org/10.3390/v14081601 - 22 Jul 2022
Cited by 9 | Viewed by 2229
Abstract
Vibrio parahaemolyticus is a major foodborne pathogen and the main cause of diarrheal diseases transmitted by seafood such as fish, shrimp, and shellfish. In the current study, a novel lytic phage infecting V. parahaemolyticus, vB_VpaP_GHSM17, was isolated from the sewage of a [...] Read more.
Vibrio parahaemolyticus is a major foodborne pathogen and the main cause of diarrheal diseases transmitted by seafood such as fish, shrimp, and shellfish. In the current study, a novel lytic phage infecting V. parahaemolyticus, vB_VpaP_GHSM17, was isolated from the sewage of a seafood market, Huangsha, Guangzhou, and its morphology, biochemistry, and taxonomy features were identified. Morphological observation revealed that GHSM17 had an icosahedral head with a short, non-contractile tail. The double-stranded DNA genome of GHSM17 consisted of 43,228 bp with a GC content of 49.42%. In total, 45 putative ORFs were identified in the GHSM17 genome. Taxonomic analysis indicated GHSM17 belonging to genus Maculvirus, family Autographiviridae. In addition, GHSM17 was stable over a wide range of temperatures (20–60 °C) and pH (5–11) and was completely inactivated after 70 min of ultraviolet irradiation. The bacterial inhibition assay revealed that GHSM17 could inhibit the growth of V. parahaemolyticus within 8 h. The results support that phage GHSM17 may be a potential candidate in the biological control of V. parahaemolyticus contamination in aquaculture. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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20 pages, 7437 KiB  
Article
Whole-Genome Analysis Reveals That Bacteriophages Promote Environmental Adaptation of Staphylococcus aureus via Gene Exchange, Acquisition, and Loss
by Wenyuan Zhou, Hua Wen, Yajie Li, Yajun Gao, Xiangfeng Zheng, Lei Yuan, Guoqiang Zhu and Zhenquan Yang
Viruses 2022, 14(6), 1199; https://doi.org/10.3390/v14061199 - 31 May 2022
Cited by 10 | Viewed by 2480
Abstract
The study of bacteriophages is experiencing a resurgence owing to their antibacterial efficacy, lack of side effects, and low production cost. Nonetheless, the interactions between Staphylococcus aureus bacteriophages and their hosts remain unexplored. In this study, whole-genome sequences of 188 S. aureus bacteriophages—20 [...] Read more.
The study of bacteriophages is experiencing a resurgence owing to their antibacterial efficacy, lack of side effects, and low production cost. Nonetheless, the interactions between Staphylococcus aureus bacteriophages and their hosts remain unexplored. In this study, whole-genome sequences of 188 S. aureus bacteriophages—20 Podoviridae, 56 Herelleviridae, and 112 Siphoviridae—were obtained from the National Center for Biotechnology Information (NCBI, USA) genome database. A phylogenetic tree was constructed to estimate their genetic relatedness using single-nucleotide polymorphism analysis. Comparative analysis was performed to investigate the structural diversity and ortholog groups in the subdividing clusters. Mosaic structures and gene content were compared in relation to phylogeny. Phylogenetic analysis revealed that the bacteriophages could be distinguished into three lineages (I–III), including nine subdividing clusters and seven singletons. The subdividing clusters shared similar mosaic structures and core ortholog clusters, including the genes involved in bacteriophage morphogenesis and DNA packaging. Notably, several functional modules of bacteriophages 187 and 2368A shared more than 95% nucleotide sequence identity with prophages in the S. aureus strain RJ1267 and the Staphylococcus pseudintermedius strain SP_11306_4, whereas other modules exhibited little nucleotide sequence similarity. Moreover, the cluster phages shared similar types of holins, lysins, and DNA packaging genes and harbored diverse genes associated with DNA replication and virulence. The data suggested that the genetic diversity of S. aureus bacteriophages was likely due to gene replacement, acquisition, and loss among staphylococcal phages, which may have crossed species barriers. Moreover, frequent module exchanges likely occurred exclusively among the subdividing cluster phages. We hypothesize that during evolution, the S. aureus phages enhanced their DNA replication in host cells and the adaptive environment of their host. Full article
(This article belongs to the Special Issue Bacteriophage Bioinformatics)
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