Mimiviridae

FAMILY MIMIVIRIDAE
Taxonomic structure of the family
Family Mimiviridae
Genus Mimivirus

Since only one genus is currently recognized, the family description corresponds to the genus
description.

GENUS MIMIVIRUS
Type species Acanthamoeba polyphaga mimivirus
Virion properties
MORPHOLOGY
Mimivirus particles are comprised of an icosahedral core protein capsid with a diameter of 0.5 μm.
This large capsid is uniformly covered with a 0.125 μm thick layer of closely packed fibers, forming
a roughly spherical object 0.75 μm in diameter that is easily visible under the light microscope with
Nomarski optics (differential interference contrast microscopy). The mimivirus particle is not entirely
symmetrical as it has a five-fold star-shaped structure at a single icosahedral vertex. This structure,
coined the stargate, extends along the whole length of the five icosahedral edges surrounding this
unique vertex (Figure 1). The central electron-dense nucleocapsid of the particle appears to be con-
tained within two 40 Å-thick lipid membranes surrounded by a 70 Å-thick protein shell (Figure 2).
PHYSICOCHEMICAL AND PHYSICAL PROPERTIES
Using ultracentrifugation in a CsCI gradient, the particle density was estimated to be about
1.36 g cm3. In addition to their unprecedented size and hairy appearance, mimivirus particles are
highly resistant to extreme conditions (pH, temperature, reducing agents) or enzymatic treatments
(proteases, glycosidases). The resilience of the particles may be due to their distinctive peripheral
fiber layer thought to be made of a dense mesh of a complex (lipo-) polysaccharide biopolymer akin
to the outer coat of bacterial spores. The presence of this outer layer, not seen in other families of
large nucleocytoplasmic DNA viruses, together with the overall size of the particle, might be linked

Figure 1: Mimivirus particle exhibiting the distinctive stargate structure (transmission electron microscopy).

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Part II – The Double Stranded DNA Viruses

Figure 2: The complex internal structure of a mimivirus particle (transmission electron microscopy).

to the heterotrophic nature of the host acanthamoeba; mimivirus particles mimic the bacteria on
which they usually feed (Figure 2).
NUCLEIC ACID
The mimivirus genome is a single linear dsDNA molecule of 1,181,549 bp with an overall nucleotide
composition of 72% (AT). Several virus-encoded mRNAs are associated with the purified particle,
but the functional significance of these findings is unknown. Mimivirus genome termini lack the
large terminal inverted repeats (up to 2 kb) found in other large DNA viruses such as phycodnavi-
ruses (chlorovirus) or poxviruses. In place of inverted terminal repeats, the mimivirus genome has
an inverted repeat of a 617-bp segment approximately 22 kb from one end of its genome and near
the other end. Pairing these regions can lead to a putative Q-like form for the mimivirus genome,
with a long (22,514 bp) and a short (259 bp) tail. The short tail does not contain any ORFs.
PROTEINS
A comprehensive proteomic study of purified virions led to the identification of the products of at least
137 mimivirus genes. Over half of these virion-associated proteins have unknown functions. The 2D
gels revealed numerous isoforms, probably due to posttranslational modifications such as glycosyla-
tion, acetylation and phosphorylation. In addition to the expected major structural components (e.g.,
the major capsid protein L425 and core L410 protein), transcription enzymes and factors (12 gene prod-
ucts) constitute the largest functional category associated with the viral particles. This set includes all
five predicted DNA-directed RNA polymerase subunits, two helicases (R350, L540), the mRNA cap-
ping enzyme and four transcription factors (L377, L538, L544, R563), including a TATA box-like binding
protein. The presence of a complete transcription machinery in mimivirus particles is reminiscent of
poxviruses. The next largest functional group contains nine gene products associated with oxidative
pathways. Protein/lipid modification enzymes are also well represented, including a phosphoesterase
and a lipase, which are eventually used for digesting the cell (vacuole) membrane, two protein kinases
and a protein phosphatase. Finally, seven proteins associated with DNA topology, damage repair and
replication are in the virion, including topoisomerases IA and IB (R194, L221), a DNA UV damage
repair endonuclease (L687) and two types (X and B) of DNA polymerases (L318, R322).
LIPIDS
The interior of the particle appears to exhibit two lipid membranes, one encircling a dark core
(nucleic acid) and one in direct contact with the interior of the protein capsid. However, this mem-
brane organization might also result from the tight folding of a single membrane. Once the infecting
virion is located within a phagocytic vacuole, the particle stargate opens allowing fusion of at least
one of the mimivirus lipid membranes with the vacuole membrane. This fusion directly delivers the
core of the particle into the host cytoplasm where it serves as a seed to initiate the formation of a

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450,000 400,000 350,000

500,000 300,000
550,000 250,000

600,000 200,000

650,000 150,000

700,000
100,000

750,000
50,000

Mimivirus
800,000 0
( 1,181,404 bp )

850,000
Cog categories

R S F Unknown
900,000
L M Q
O E N
950,000 K J Other

1,000,000

1,050,000
1,100,000
1,150,000

Figure 3: Map of the mimivirus chromosome. The predicted protein coding sequences are shown on both
strands and colored according to the function category of their matching COG. Genes with no COG match are
shown in gray. Abbreviations for the COG functional categories are as follows: E, amino acid transport and
metabolism; F, nucleotide transport and metabolism; J, translation; K, transcription; L, replication, recombina-
tion, and repair; M, cell wall/membrane biogenesis; N, cell motility; O, posttranslational modification, protein
turnover, and chaperones; Q, secondary metabolites biosynthesis, transport, and catabolism; R, general func-
tion prediction only; S, function unknown. Small red arrows indicate the location and orientation of tRNAs.
The AC excess profile is shown on the innermost circle, exhibiting a peak around position 380,000. (From
Raoult et al. (2004). Science, 306, 1344–1350; with permission of AAAS.)

virion factory. As expected from the crucial role likely to be played by the internal lipid membranes
of the particle in the infection process, mimivirus virions are readily inactivated by treatment with
lipophilic compounds (such as DMSO).
CARBOHYDRATES
The ultra-structure of the mature particle (as observed by electron or atomic-force microscopy) as
well as preliminary biochemical and mass spectrometry analyses suggest that the outer fiber layer
might consist of a biopolymer similar to peptidoglycan. Accordingly, the mimivirus genome encodes
a number of proteins homologous to enzymes key to the synthesis of bacterial cell walls. Among
those proteins, the functions of a UDP-D-glucose 4,6-dehydratase (R141) and of a bifunctional UDP-
4-keto-6-deoxy-D-glucose epimerase/reductase (L780) have been experimentally validated.

Genome organization and replication

The initial mimivirus genome annotation predicted 911 protein-coding genes and 6 tRNAs (Figure 3).
More recent data obtained through transcriptome sequencing (RNA-Seq) and deep genome

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resequencing allowed the identification of a total of 1018 genes, including 979 protein-coding genes,
6tRNAs and 33 non-coding mRNAs. The latest genome sequence and the most current annotation
(including the location of identified promoter signals and known 5-end and 3-end transcript bound-
aries) is available in the RefSeq database under accession number NC_014649.1, and in GenBank
under accession number HQ336222.

The penetration of the particle inner core within the host cytoplasm is followed by a complete eclipse
phase that lasts approximately two hours in Acanthamoeba castellanii (ATCC 30010), after which time
mimivirus virion factories become visible. Mimivirus replication entirely takes place in the cyto-
plasm of the host Acanthamoeba cell, through the successive expression of early (from 0 to 3 h post
infection), intermediate (from 3 h to 6 h post-infection) and late (after 6 h post-infection) transcripts,
each gene class representing approximately one-third of the mimivirus genome. The virion factories
develop from the core of individual uncoated virus particles (seeds). The earliest viral transcripts are
detected as soon as 15 minutes post infection, most likely produced by the viral transcription machin-
ery within the uncoated particles. Most of the genes involved in nucleotide synthesis and DNA
replication are transcribed from 3 h to 6 h post-infection. Late genes (after 6 h) include virion struc-
tural components, as well as most of the virally-encoded transcription apparatus components. This
expression pattern suggests that the early and intermediate mimivirus transcripts detected before the
appearance of fully mature cytoplasmic virion factories are generated by the transcription apparatus
associated with the virion core. Mimivirus particles (at least one thousand per infected cell) are con-
tinually produced for up to 12 h by the growing virion factories (up to 6 μm in diameter) (Figure 4).
Mature mimivirus particles increasingly fill the host cytoplasm and are progressively released from
the dying cell. No budding or sudden cell bursts are seen.

Figure 4: The distinctive giant mimivirus virion factory in full production (8 h post infection in Acanthamoeba
castellanii). The dark circle (about 4.5 μm in diameter) is the virion factory from which mimivirus particles can
be seen emerging, first empty, then filled with a dense core, then covered with their outer fiber layer (transmis-
sion electron microscopy).

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Antigenic properties and pathogenicity

Acanthamoeba polyphaga mimivirus was isolated from the water of a cooling tower in Bradford,
England. Mimivirus readily infects many Acanthamoeba strains, including its preferred laboratory
host Acanthamoeba castellanii. Metagenomic surveys indicate that close relatives of the Mimiviridae
family are prevalent in the sea, where they probably infect marine heterotrophic protists and regu-
late plankton populations.

Although mimivirus was isolated in the context of a pneumonia epidemic and initially thought to
be an emerging human pathogen based on positive serology, subsequent more specific PCR-based
studies failed to detect mimivirus in large numbers of pneumonia patients. As expected from the
size of its particle, mimivirus is internalized by various professional phagocytic cells, including
human macrophages in vitro. However, the recent demonstration that mimivirus particles are rec-
ognized by the sera of patients infected by Francisella tularensis, casts serious doubt on previously
published serological evidence for mimivirus infection of humans. At the moment, there is little evi-
dence that mimivirus is a human pathogen.

Biological properties
The presence of many proteins never seen in any other virus is one of the unique features of mimivi-
rus. In addition to the full DNA replication and transcription apparatus usually found in large eukaryo-
tic DNA viruses (such as the poxviruses), mimivirus encodes many enzymes central to the translation
apparatus such as four aminoacyl-tRNA synthase: Tyr-RS, Met-RS, Arg-RS and Cys-RS. Mimivirus
also possesses the first virally-encoded nucleotide diphosphate kinase (NDK), mismatch repair
MutS-homolog, and tRNA (Uracil-5-)-methyltransferase. The mimivirus genome also encodes sev-
eral sugar-manipulating enzymes likely to be involved in the biosynthesis of the particle outer layer.
Crystallographic 3D structures have been obtained for the following mimivirus proteins: Tyr-RS (L124),
NDK (R418), formamidopyrimidine-DNA glycosylase (L315), sulfhydryl oxidase (R596), mRNA-cap-
ping enzyme (R382), and mimivirus cyclophilin (L605). A new type of satellite virus (called the Sputnik
virophage) has been isolated in association with a new strain of mimivirus (called mamavirus).

Species demarcation criteria in the genus

Not applicable.

List of species in the genus Mimivirus

Acanthamoeba polyphaga mimivirus
Acanthamoeba polyphaga mimivirus [HQ336222  NC_014649] (APMV)
(Rowbotham-Bradford)
Acanthamoeba polyphaga mamavirus (La Scola-Paris) [EU827539, EU827540, EU827541] (APMV2)
Megavirus chilensis
Megavirus chilensis (Claverie-Las Cruces) [JN258408] (McV)
Species names are in italic script; names of isolates are in roman script. Sequence accession numbers [ ] and assigned
abbreviations ( ) are also listed.

List of other related viruses which may be members of the genus Mimivirus but have not
been approved as species
Terravirus
Courdovirus
Moumouvirus

Derivation of name
Mimi: for mimicking microbe.

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Further reading
Journals and books
Abergel, C., Rudinger-Thirion, J., Giegé, R. and Claverie, J.M. (2007). Virus-encoded aminoacyl-tRNA synthetases: structural
and functional characterization of mimivirus TyrRS and MetRS. J. Virol., 81, 12406–12417.
Claverie, J.M. and Abergel, C. (2009). Mimivirus and its virophage. Annu. Rev. Genet., 43, 49–66.
La Scola, B., Desnues, C., Pagnier, I., Robert, C., Barrassi, L., Fournous, G., Merchat, M., Suzan-Monti, M., Forterre, P.,
Koonin, E. and Raoult, D. (2008). The virophage as a unique parasite of the giant Mimivirus. Nature, 455, 100–104.
Legendre, M., Audic, S., Poirot, O., Byrne, D., Lartigue, A., Lescot, M., Bernadac, A., Poulain, J., Abergel, C. and Claverie,
J.M. (2010). mRNA deep sequencing reveals 75 new genes and a complex transcriptional landscape in Mimivirus.
Genome Res., 20, 664–674.
Monier, A., Claverie, J.M. and Ogata, H. (2008). Taxonomic distribution of large DNA viruses in the sea. Genome Biol., 9, R106.
Mutsafi, Y., Zauberman, N., Sabanay, I. and Minsky, A. (2010). Vaccinia-like cytoplasmic replication of the giant Mimivirus.
Proc. Natl Acad. Sci., U S A, 107, 5978–5982.
Pelletier, N., Raoult, D. and La Scola, B. (2009). Specific recognition of the major capsid protein of Acanthamoeba polyphaga
Mimivirus by sera of patients infected by Francisella tularensis. FEMS Microbiol Lett., 297, 117–123.
Raoult, D., Audic, S., Robert, C., Abergel, C., Renesto, P., Ogata, H., La Scola, B., Suzan, M. and Claverie, J.M. (2004). The
1.2-Mb genome sequence of mimivirus. Science, 306, 1344–1350.
Xiao, C., Kuznetsov, Y.G., Sun, S., Hafenstein, S.L., Kostyuchenko, V.A., Chipman, P.R., Suzan-Monti, M., Raoult, D.,
McPherson, A. and Rossmann, M.G. (2009). Structural studies of the giant mimivirus. PLoS Biol., 7, e92.
Zauberman, N., Mutsafi, Y., Halevy, D.B., Shimoni, E., Klein, E., Xiao, C., Sun, S. and Minsky, A. (2008). Distinct DNA exit
and packaging portals in the virus Acanthamoeba polyphaga mimivirus. PLoS Biol., 6, e114.

Websites
Results for Mimivirus at http://www.igs.cnrs-mrs.fr

Contributed by
Claverie, J-M. and Abergel, C.

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