Phage Therapy: Beyond Antibacterial Action.

Until recently, phages were considered as mere "bacteria eaters" with potential for use in combating antimicrobial resistance. The real value of phage therapy assessed according to the standards of evidence-based medicine awaits confirmation by clinical trials. However, the progress in research on phage biology has shed more light on the significance of phages. Accumulating data indicate that phages may also interact with eukaryotic cells. How such interactions could be translated into advances in medicine (especially novel means of therapy) is discussed herein.

Complete Genome Sequences of Two Novel Staphylococcus aureus Podoviruses of Potential Therapeutic Use, vB_SauP_phiAGO1.3 and vB_SauP_phiAGO1.9.

Here, we report the genome sequences of two Staphylococcus aureus phages belonging to the family Podoviridae and subfamily Picovirinae, vB_SauP_phiAGO1.3 and vB_SauP_phiAGO1.9, which were isolated from Warsaw sewage. Analysis of their genomes provides valuable information about the diversity of phages belonging to the genus Rosenblumvirus and their genes that undergo evolutionary adaptation to cells of different host strains.

Methods for Bacteriophage Preservation.

In a view of growing interest in bacteriophages as the most abundant members of microbial communities and as antibacterial agents, reliable methods for bacteriophage long-term preservation, that warrant the access to original or mutant stocks of unchanged properties, have become of crucial importance. A storage method that retains the infectivity of any kind of bacteriophage virions, either in a cell lysate or in a purified suspension, does not exist, due to the enormous diversity of bacteriophages and hence the differentiation of their sensitivity to various storage conditions. Here, we describe a method of long-term bacteriophage preservation, which is based on freezing of freshly infected susceptible bacteria at early stages of bacteriophage development. The infected bacteria release mature bacteriophages upon melting enabling the recovery of bacteriophage virions with high efficiency. The only limitation of this method is the sensitivity of bacteriophage host to deep-freezing, and thus it can be used for the long-term preservation of the vast majority of bacteriophages.

Corrigendum: Bacteriophage Procurement for Therapeutic Purposes.

[This corrects the article on p. 1177 in vol. 7, PMID: 27570518.].

[Lysis of bacterial cells in the process of bacteriophage release--canonical and newly discovered mechanisms]. / Liza komórek bakteryjnych w procesie uwalniania bakteriofagów ­ kanoniczne i nowo poznane mechanizmy.

The release of phage progeny from an infected bacterium is necessary for the spread of infection. Only helical phages are secreted from a cell without causing its destruction. The release of remaining phages is correlated with bacterial lysis and death. Thus, the understanding of phage lytic functions is crucial for their use in the fight with bacterial pathogens. Bacteriophages with small RNA or DNA genomes encode single proteins which are called amurins and cause lysis by the inhibition of cell wall synthesis. Bacteriophages of double-stranded DNA genomes, which dominate in the environment, encode enzymes that are called endolysins and contribute to lysis by the cleavage of cell wall peptydoglycan. Endolysins that do not contain signal sequences cannot pass the cytoplasmic membrane by themselves. Their access to peptidoglycan is provided by membrane proteins - holins, which can form in the membrane large pores, that are called "holes". Some endolysins do not require holins for their transport, owing to the presence of the so called SAR sequence at their N-terminus. It enables their transport through the membrane by the bacterial sec system. However, it is not cleaved off, and thus these endolysins remain trapped in the membrane in an inactive form. Their release, which is correlated with the activation, occurs as a result of membrane depolarization and depends on proteins that are called pinholins. Pinholins form in membrane pores that are too small for the passage of endolysins but sufficient for membrane depolarization. Proteins that are called antiholins regulate the timing of lysis, through the blockage of holins action until the end of phage morphogenesis. Additionally, newly identified lytic proteins, spanins, participate in the release of progeny phages from Gram-negative bacteria cells. They cause the destruction of outer cell membrane by its spanning with the cytoplasmic membrane. This is possible after the endolysin-mediated destruction of peptidoglycan, which separates both membranes, and ensures the fast completion of lysis.

A reliable method for storage of tailed phages.

A universal and effective method for long-term storage of bacteriophages has not yet been described. We show that randomly selected tailed phages could be stored inside the infected cells at -80°C without a major loss of phage and host viability. Our results suggest the suitability of this method as a standard for phage preservation.

Genome of bacteriophage P1.

P1 is a bacteriophage of Escherichia coli and other enteric bacteria. It lysogenizes its hosts as a circular, low-copy-number plasmid. We have determined the complete nucleotide sequences of two strains of a P1 thermoinducible mutant, P1 c1-100. The P1 genome (93,601 bp) contains at least 117 genes, of which almost two-thirds had not been sequenced previously and 49 have no homologs in other organisms. Protein-coding genes occupy 92% of the genome and are organized in 45 operons, of which four are decisive for the choice between lysis and lysogeny. Four others ensure plasmid maintenance. The majority of the remaining 37 operons are involved in lytic development. Seventeen operons are transcribed from sigma(70) promoters directly controlled by the master phage repressor C1. Late operons are transcribed from promoters recognized by the E. coli RNA polymerase holoenzyme in the presence of the Lpa protein, the product of a C1-controlled P1 gene. Three species of P1-encoded tRNAs provide differential controls of translation, and a P1-encoded DNA methyltransferase with putative bifunctionality influences transcription, replication, and DNA packaging. The genome is particularly rich in Chi recombinogenic sites. The base content and distribution in P1 DNA indicate that replication of P1 from its plasmid origin had more impact on the base compositional asymmetries of the P1 genome than replication from the lytic origin of replication.