Kinetoplast

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Electron micrograph of normal kinetoplast (K) of Trypanosoma brucei Kinetoplast of Trypanosoma brucei.tif
Electron micrograph of normal kinetoplast (K) of Trypanosoma brucei

A kinetoplast is a network of circular DNA (called kDNA) inside a mitochondrion that contains many copies of the mitochondrial genome. [1] [2] The most common kinetoplast structure is a disk, but they have been observed in other arrangements. Kinetoplasts are only found in Excavata of the class Kinetoplastida. The variation in the structures of kinetoplasts may reflect phylogenic relationships between kinetoplastids. [3] A kinetoplast is usually adjacent to the organism's flagellar basal body, suggesting that it is bound to some components of the cytoskeleton. In Trypanosoma brucei this cytoskeletal connection is called the tripartite attachment complex and includes the protein p166. [4]

Contents

Trypanosoma

In trypanosomes, a group of flagellated protozoans, the kinetoplast exists as a dense granule of DNA within the mitochondrion. Trypanosoma brucei , the parasite which causes African trypanosomiasis (African sleeping sickness), is an example of a trypanosome with a kinetoplast. Its kinetoplast is easily visible in samples stained with DAPI, a fluorescent DNA stain, or by the use of fluorescent in situ hybridization (FISH) with BrdU, a thymidine analogue. [5]

Structure

The kinetoplast contains circular DNA in two forms, maxicircles and minicircles. Maxicircles are between 20 and 40kb in size and there are a few dozen per kinetoplast. There are several thousand minicircles per kinetoplast and they are between 0.5 and 1kb in size. Maxicircles encode the typical protein products needed for the mitochondria which is encrypted. Herein lies the only known function of the minicircles - producing guide RNA (gRNA) to decode this encrypted maxicircle information, typically through the insertion or deletion of uridine residues. The network of maxicircles and minicircles are catenated to form a planar network that resembles chain mail. Reproduction of this network then requires that these rings be disconnected from the parental kinetoplast and subsequently reconnected in the daughter kinetoplast. [5] [6] This unique mode of DNA replication may inspire potential drug targets.

The best studied kDNA structure is that of Crithidia fasciculata , a catenated disk of circular kDNA maxicircles and minicircles, most of which are not supercoiled. [3] Exterior to the kDNA disk but directly adjacent are two complexes of proteins situated 180˚ from each other and are involved in minicircle replication. [1] [2] [5] [6]

Variations

Variations of kinetoplast networks have also been observed and are described by the arrangement and location of their kDNA.

The presence of this variety of kDNA structures reinforces the evolutionary relationship between the species of kinetoplastids. As pan-kDNA most closely resembles a DNA plasmid, it may be the ancestral form of kDNA. [3]

Replication

Figure 8. Illustration of the location of the antipodal protein complex relative to kinetoplast disk (above) and the migration of minicircle to these complexes for replication (below). Kinetoplast Replication.jpg
Figure 8. Illustration of the location of the antipodal protein complex relative to kinetoplast disk (above) and the migration of minicircle to these complexes for replication (below).

The replication of the kinetoplast occurs simultaneously to the duplication of the adjacent flagellum and just prior to the nuclear DNA replication. In a traditional Crithidia fasciculata kDNA network, initiation of replication is promoted by the unlinking of kDNA minicircles via topoisomerase II. The free minicircles are released into a region between the kinetoplast and the mitochondrial membrane called the kinetoflagellar zone (KFZ). [2] [3] [6] After replication the minicircles migrate by unknown mechanisms to the antipodal protein complexes that contain several replication proteins including an endonuclease, helicase, DNA polymerase, DNA primase, and DNA ligase, which initiate repair of remaining discontinuities in the newly replicated minicircles. [5]

This process occurs one minicircle at a time, and only a small number of minicircles are unlinked at any given moment. To keep track of which minicircles have been replicated, upon rejoining to the kDNA network a small gap remains in the nascent minicircles, which identifies them as having already been replicated. Minicircles that have not yet been replicated are still covalently closed. Immediately after replication, each progeny is attached to the kDNA network proximal to the antipodal protein complexes and the gaps are partially repaired. [1] [6]

Figure 9. Illustration of kinetoplast rotation during minicircle replication. Kinetoplast Rotation.jpg
Figure 9. Illustration of kinetoplast rotation during minicircle replication.
Kinetoplast (K) divides first and then the nucleus (N) in dividing T. brucei Kinetoplast dividing.TIF
Kinetoplast (K) divides first and then the nucleus (N) in dividing T. brucei

As minicircle replication progresses, to prevent the build-up of new minicircles, the entire kDNA network will rotate around the central axis of the disk. The rotation is believed to be directly connected to the replication of the adjacent flagellum, as the daughter basal body will also rotate around the mother basal body in a timing and manner similar to the rotation of the kinetoplast. By rotating, the minicircles of the daughter kinetoplast are assembled in a spiral fashion and begin moving inward toward the center of the disk as new minicircles are unlinked and moved into the KFZ for replication. [2] [5] [6]

While the exact mechanisms for maxicircle kDNA have yet to be determined in the same detail as minicircle kDNA, a structure called a nabelschnur (German for "umbilical cord") is observed that tethers the daughter kDNA networks but eventually breaks during separation. Using FISH probes to target the nabelschnur, it has been found to contain maxicircle kDNA. [5]

Kinetoplast replication is described as occurring in five stages, each in relation to the replication of the adjacent flagellum.

DNA repair

Trypanosoma cruzi is able to repair nucleotides in its genomic or kinetoplast DNA that have been damaged by reactive oxygen species produced by the parasite's host during infection. [7] DNA polymerase beta expressed in T. cruzi is employed in the removal of oxidative DNA damages by the process of base excision repair. It appears that DNA polymerase beta acts during kinetoplast DNA replication to repair oxidative DNA damages induced by genotoxic stress in this organelle. [7]

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<span class="mw-page-title-main">Trypanosomatida</span> Order of flagellate protists in the kinetoplastid excavates

Trypanosomatida is a group of kinetoplastid unicellular organisms distinguished by having only a single flagellum. The name is derived from the Greek trypano (borer) and soma (body) because of the corkscrew-like motion of some trypanosomatid species. All members are exclusively parasitic, found primarily in insects. A few genera have life-cycles involving a secondary host, which may be a vertebrate, invertebrate or plant. These include several species that cause major diseases in humans. Some trypanosomatida are intracellular parasites, with the important exception of Trypanosoma brucei.

<span class="mw-page-title-main">Kinetoplastida</span> Flagellated protists belonging to the phylum Euglenozoa

Kinetoplastida is a group of flagellated protists belonging to the phylum Euglenozoa, and characterised by the presence of an organelle with a large massed DNA called kinetoplast. The organisms are commonly referred to as "kinetoplastids" or "kinetoplasts" The group includes a number of parasites responsible for serious diseases in humans and other animals, as well as various forms found in soil and aquatic environments. Their distinguishing feature, the presence of a kinetoplast, is an unusual DNA-containing granule located within the single mitochondrion associated with the base of the cell's flagellum. The kinetoplast contains many copies of the mitochondrial genome.

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Trypanosoma brucei is a species of parasitic kinetoplastid belonging to the genus Trypanosoma that is present in sub-Saharan Africa. Unlike other protozoan parasites that normally infect blood and tissue cells, it is exclusively extracellular and inhabits the blood plasma and body fluids. It causes deadly vector-borne diseases: African trypanosomiasis or sleeping sickness in humans, and animal trypanosomiasis or nagana in cattle and horses. It is a species complex grouped into three subspecies: T. b. brucei, T. b. gambiense and T. b. rhodesiense. The first is a parasite of non-human mammals and causes nagana, while the latter two are zoonotic infecting both humans and animals and cause African trypanosomiasis.

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References

  1. 1 2 3 Shapiro TA; Englund PT (1995). "The structure and replication of kinetoplast DNA". Annu. Rev. Microbiol. 49: 117–43. doi:10.1146/annurev.mi.49.100195.001001. PMID   8561456.
  2. 1 2 3 4 Shlomai J (2004). "The structure and replication of kinetoplast DNA". Curr. Mol. Med. 4 (6): 623–47. doi:10.2174/1566524043360096. PMID   15357213.
  3. 1 2 3 4 5 6 7 8 Lukes J, et al. (2002). "Kinetoplast DNA Network: Evolution of an Improbable Structure". Eukaryotic Cell. 1 (4): 495–502. doi:10.1128/ec.1.4.495-502.2002. PMC   117999 . PMID   12455998.
  4. Zhao, Z; Lindsay, M. E.; Roy Chowdhury, A; Robinson, D. R.; Englund, P. T. (2008). "P166, a link between the trypanosome mitochondrial DNA and flagellum, mediates genome segregation". The EMBO Journal. 27 (1): 143–54. doi:10.1038/sj.emboj.7601956. PMC   2206137 . PMID   18059470.
  5. 1 2 3 4 5 6 7 Gluenz E, et al. (March 2011). "The kinetoplast replication cycle in Trypanosoma brucei is orchestrated by cytoskeleton-mediated cell morphogenesis". Molecular Cell Biology. 31 (5): 1012–1021. doi:10.1128/MCB.01176-10. PMC   3067821 . PMID   21173163.
  6. 1 2 3 4 5 Torri, A., et al.DNA Replication in Eukaryotic Cells. Cold Spring Harbor Laboratory Press. 1996. pages=1029–42. ISBN   0-87969-459-9
  7. 1 2 Schamber-Reis BL, Nardelli S, Régis-Silva CG, Campos PC, Cerqueira PG, Lima SA, Franco GR, Macedo AM, Pena SD, Cazaux C, Hoffmann JS, Motta MC, Schenkman S, Teixeira SM, Machado CR (2012). "DNA polymerase beta from Trypanosoma cruzi is involved in kinetoplast DNA replication and repair of oxidative lesions". Mol. Biochem. Parasitol. 183 (2): 122–31. doi: 10.1016/j.molbiopara.2012.02.007 . PMID   22369885.