Filamentous bacteriophage

The filamentous particle seen in electron micrographs was initially interpreted as contaminating pili, but ultrasonic degradation, which breaks flexible filaments roughly in half,[5] inactivated infectivity as predicted for a filamentous phage morphology.[6] Three filamentous bacteriophages, fd, f1 and M13, were isolated and characterized by three different research groups in the early 1960s. Since these three phages differ by less than 2 percent in their DNA sequences, corresponding to changes in only a few dozen codons in the whole genome, for many purposes they can be considered to be identical. Further independent characterization was shaped by the interests of the separate research groups.[2] Filamentous phages, unlike most other phages, are continually extruded through the bacterial membrane without killing the host.[7] Genetic studies on M13 using conditional lethal mutants, initiated by David Pratt and colleagues, led to description of phage gene functions.[8][9] Notably, the protein product of gene 5, which is required for synthesis of progeny single-stranded DNA, is made in large amounts in the infected bacteria,[10][11][12] and it binds to the nascent DNA to form a linear intracellular complex.[13] (The simple numbering of genes using Arabic numerals 1,2,3,4… introduced by the Pratt group is sometimes displaced by the practice, introduced by some f1 researchers, of using Roman numerals I, II, III, IV… but the gene numbers defined by the two systems are the same).

Longer (or shorter) DNA can be included in fd phage, since more (or fewer) protein subunits can be added during assembly as required to protect the DNA, making the phage convenient for genetic studies.[14] The length of the phage is also affected by the positive charge per length on the inside surface of the phage capsid.[15] M13 is widely used for research involving phage mutants, and is sometimes called the type species. The genome of fd was one of the first complete genomes to be sequenced.[16]

The taxonomy of filamentous bacteriophage was defined by Andre Lwoff and Paul Tournier as family Inophagoviridae, genus I. inophagovirus, species Inophagovirus bacterii (Inos=fiber or filament in Greek), with phage fd (Hoffmann-Berling) as the type species.[17] However, "Phagovirus" is tautological, and the name of the family was altered to Inoviridae and the type genus to Inovirus. This nomenclature persisted for many decades, but the number of known filamentous bacteriophages has multiplied many-fold by using a machine-learning approach, and “the former Inoviridae family should be reclassified as an order, provisionally divided into 6 candidate families and 212 candidate subfamilies”.[18] Phages fd, f1, M13 and other related phages are often referred to as members of the Ff group of phages, for F specific (they infect Escherichia coli carrying the F-episome) filamentous phages, using the concept of vernacular name.[19]

George Smith and Greg Winter used f1 and fd for their work on phage display for which they were awarded a share of the 2018 Nobel Prize in Chemistry.[20] The creation and exploitation of many derivatives of M13 for a wide range of purposes, especially in materials science, has been employed by Angela Belcher and colleagues.[21] Filamentous bacteriophage can promote antibiotic tolerance by forming liquid crystalline domains around bacterial cells.[22].[23]

The molecular structure of Ff filamentous phage was determined using a number of physical techniques, especially X-ray fiber diffraction.[2][24] Filamentous phage were further refined using solid-state NMR and cryo-electron microscopy.[2][23] The single-stranded Ff phage DNA runs down the central core of the phage, and is protected by a cylindrical protein coat built from thousands of identical α-helical protein subunits coded by phage gene 8. The gene 8 protein is inserted in the plasma membrane as an early step in phage assembly.[2] Some strains of phage have a "leader sequence" to promote membrane insertion, but others do not seem to need the leader sequence. The two ends of the phage are capped by a few copies of proteins that are important for infection of the host bacteria, and also for assembly of nascent phage particles. These proteins are the products of phage genes 3 and 6 at one end of the phage, and phage genes 7 and 9 at the other end. The fiber diffraction studies identified two structural classes of phage, differing in the details of the arrangement of the gene 8 protein. Class I, including strains fd, f1, M13, If1 and IKe, has a rotation axis relating the gene 8 coat proteins, whereas Class II, including strains Pf1, Pf3, Pf4 and PH75, this rotation axis is replaced by a helix axis. This technical difference has little noticeable effect on the overall phage structure, but the extent of independent diffraction data is greater for symmetry Class II than for Class I. This assisted the determination of the Class II phage Pf1 structure, and by extension the Class I structure.[24]

The DNA isolated from fd phage is single-stranded, and topologically a circle. That is, the DNA single strand extends from one end of the phage particle to the other and then back again to close the circle, although the two strands are not base-paired. This topology was assumed to extend to all other filamentous phages, but it is not the case for phage Pf4, for which the DNA in the phage is topologically linear, not circular.[23] During fd phage assembly, the phage DNA is first packaged in a linear intracellular nucleoprotein complex with many copies of the phage gene 5 replication/assembly protein, which is displaced by the gene 8 coat protein as the nascent phage is extruded across the bacterial plasma membrane without killing the bacterial host.[13][25][2][7] This assembly mechanism makes this phage a valuable system with which to study transmembrane proteins.[2][4]

• Ff phages – these infect Escherichia coli carrying the F episome.
  • M13 bacteriophage
  • f1 phage
  • fd phage
• Pf1 phage —  bacteriophage that infects Pseudomonas aeruginosa

Filamentous bacteriophages are classified in the family Inoviridae, genus inovirus.[26] Phylogenetic trees show the relations between different strains of inovirus.[1][3][18]