| Microviridae | |
|---|---|
Virus classification | |
| (unranked): | Virus |
| Realm: | Monodnaviria |
| Kingdom: | Sangervirae |
| Phylum: | Phixviricota |
| Class: | Malgrandaviricetes |
| Order: | Petitvirales |
| Family: | Microviridae |
| Subfamilies and genera | |
Subfamily:Bullavirinae Subfamily:Gokushovirinae | |
Microviridae is a family ofbacteriophages with a single-stranded DNA genome. The name of this family is derived from theancient Greek wordμικρός (mikrós), meaning "small".[1][2] This refers to the size of their genomes, which are among the smallest of the DNA viruses.Enterobacteria, intracellular parasitic bacteria, andspiroplasma serve as natural hosts. There are 22 species in this family, divided among seven genera and two subfamilies.[3][4]
The virions are non-enveloped, round with an icosahedral symmetry (T = 1). They have a diameter between 25 and 27nanometers and lack tails. Each virion has 60 copies each of the F, G, and J proteins and 12 copies of the H protein. They have 12 pentagonal trumpet-shaped pentamers (~7.1 nm wide × 3.8 nm high), each of which is composed of 5 copies of the G and one of the H protein.[citation needed]
Viruses in this family replicate their genomes via a rolling circle mechanism and encode dedicated RCR initiation proteins.[5][6]
Although the majority of species in this family have lytic life cycles, a few may have temperate life cycles.[7]
The genome sizes range from 4.5–6kb and is circular. It encodes 11genes (in order: A, A*, B, C, K, D, E, J, F, G, and H), nine of which are essential. The nonessential genes are E and K.[8][9] Several of the genes have overlapping reading frames.[10][11] Protein A* is encoded within protein A. It lacks ~1/3 of the amino acids from the N terminal of the A protein and is encoded in the same frame as the A protein. It is translated from an internal start site within the messenger RNA. Gene E is encoded with gene D with a +1 frameshift. Gene K overlaps genes A, B, and C. The origin of replication lies within a 30 base sequence.[12] The entire 30 base sequence is required for replication.[13]
The majorcapsid protein (F) has 426amino acids, the majorspike protein (G) has 175 amino acids, the small DNA-binding protein (J) has 25–40 amino acids, and the DNApilot protein (H) has 328 amino acids.[14] The major folding motif of protein F is the eight-stranded antiparallel beta barrel common to many viral capsid proteins.[15] The G protein is a tight beta barrel with its strands running radially outward. The G proteins occur in groups of five forming 12 spikes that enclose a hydrophilic channel. The highly basic J protein lacks any secondary structure and is situated in an interior cleft of the F protein. It has no acidic amino acid residues in the protein and the twelve basic residues are concentrated in two clusters in the N-terminus separated by a proline-rich region.[citation needed]
Assembly of the virion uses two scaffolding proteins, internal scaffolding protein B and external scaffolding protein D. The function of protein B seems to be to lower the amount of protein D needed by the virion for assembly.[16] Protein H is a multifunctional structural protein required for piloting the viral DNA into the host cell interior during the entry process. Protein E is a 91-amino acid membrane protein that causes host cell lysis by inhibiting the hosttranslocase MraY.[17] This inhibitory activity is located within the N terminal 29 amino acids.[18] Protein A is a single strandendonuclease[19] and is responsible for the initiation of viral DNA replication.[20] It catalyses cleavage and ligation of a phosphodiester bond between aG andA nucleotide residue pair at the phi X origin.[21] It may not be essential for phage viability but burst sizes are reduced by 50% when it is mutated.[22] Protein A* inhibits host DNA replication.[23] Unlike protein A it is capable of cleaving the phi X viral DNA in the presence of single-stranded binding protein of the host.[24] Protein A*, like Protein A, may not be required for phage viability.[25] Protein C increases the fidelity of the termination and reinitiation reactions and is required for the packagaging of the viral DNA in to the protein shell.[26] Protein K has 56 amino acids and is found in the membrane of the host cell. It appears to be able to increase the burst size of the virus.[27]
This family is divided into two subfamilies:Gokushovirinae andBullavirinae (former genusMicrovirus). These groups differ in their hosts, genome structure, and viron composition. The nameGokushovirinae is derived from theJapanese forvery small. Gokushoviruses are currently known to infect only obligate intra-cellular parasites. The members of the subfamilyBullavirinae all infectEnterobacteria.[citation needed]
A putative third grouping has been proposed—Alpavirinae—which infect the orderBacteroidales.[7] This a group of viruses known only asprophages and additional work on these viruses seems indicated before subfamily status is granted.
A fourth clade has been proposed—Pichovirinae.[28] This clade has a genome organisation that differs from the other members of this family. The name is derived frompicho which meanssmall inOccitan.[citation needed]
Another virus has been isolate from the turkey gut with features similar to other microviruses but quite distinct from the known species.[29]
Members of the subfamilyBullavirinae (former genusMicrovirus) have four structural proteins: major capsid protein F, major spike protein G, a small DNA-binding protein J (25 - 40 amino acids in length) and DNA pilot protein H. Assembly of the viron uses two scaffolding proteins, internal scaffolding protein B and external scaffolding protein D. Protein H is a multifunctional structural protein required for piloting the viral DNA into the host cell interior during the entry process. The genomes are between 5.3 and 6.2 kilobases (kb) in length.[citation needed]
Members of this subfamily can be separated into three mainclades according to genome sizes.[30] Size variability within the groups occurs mainly as a result of insertions and deletions of the intergenic regions. Viruses are assigned according to their similarity to known lab based strains—the ΦX174-like clade, G4-like clade and the α3-like clade. The ΦX174-like clade of microviridae have the smallest and least variable genomes (5,386–5,387 bp); the G4-like clade varies in size from 5,486 to 5,487 bp; while the largest genome sized group is the α3-like clade with genomes ranging from 6,061–6259bp.[citation needed]
Members of the subfamilyGokushovirinae have only two structural proteins: capsid proteins F (Virus Protein 1) and DNA pilot protein H (Virus Protein 2) and do not use scaffolding proteins. They also possess 'mushroom-like' protrusions positioned at the three-fold axes of symmetry of their icosahedral capsids. These are formed by large insertion loops within the protein F of gokushoviruses and are absent in the microviruses. They lack both the external scaffolding protein D and the major spike protein G of the species in the genusMicrovirus. The genomes of this group tend to be smaller—about 4.5 kb in length. This subfamily includes the generaBdellomicrovirus,Chlamydiamicrovirus andSpiromicrovirus.[citation needed]
There are a number of steps in the life cycle
1. Adsorption to the host via specific receptor(s)
2. Movement of the viral DNA into the host cell
3. Conversion of the single strand form to a double-stranded intermediate
This is known as the replicative form I.
4. Transcription of early genes
5. Replication of the viral genome
Viral protein A cleaves replicative form I DNA strand at the origin of replication (ori) and covalently attaches itself to the DNA, generating replicative form II molecule. Replication of the genome now begins via a rolling circle mechanism. The host'sDNA polymerase converts the single-stranded DNA into double-stranded DNA.
6. Late genes are now transcribed by the host'sRNA polymerase.
7. Synthesis of the new virons
Viral protein C binds to replication complex, inducing packaging of new viral positive-stranded DNA intoprocapsids.[31] The preinitiation complex consists of the host cell proteinrep and viral A and C proteins.[32] These associate with the procapsid forming a 50S complex.
8. Maturation of the virons in the host cytoplasm
9. Release from the host
Celllysis is mediated by thephiX174-encoded protein E, which inhibits thepeptidoglycan synthesis leading to an eventual bursting of the infected cell.[citation needed]
Roykta, D.R. et al., 2006. Horizontal Gene Transfer and the Evolution of MicroviridColiphage Genomes. Journal of Bacteriology, 118(3) p1134–1142
