Functional Identification of the Plasmodium Centromere and Generation of a Plasmodium Artificial Chromosome

Iwanaga, Shiro; Khan, Shahid M.; Kaneko, Ikuyo; Christodoulou, Zóe; Newbold, Chris; Yuda, Morihide; Janse, Chris J.; Waters, Andy

doi:10.1016/j.chom.2010.04.011

Cited by 14 publications

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“…This sequence variability precludes categorization as point centromeres, and the presence of a single peak (with rare exceptions, described below) rather than recruitment of CENH3 across the entire chromosome indicates the absence of a holocentric chromosome. The regional centromere structure is found in most eukaryotes, including the closest related organisms with identified centromeres: the protist Plasmodium falciparum and the red alga C. merolae (21)(22)(23)(24). Both organisms have similarly sized centromeric DNA regions (∼2-4 kb) and also share low-GC content as a characteristic of their centromeres: ∼3% relative to the genome average GC of 21.8% in the case of P. falciparum (21,22), and 48.4% relative to the genome average GC of 55% for C. merolae (24).…”

Section: Discussionmentioning

confidence: 99%

“…The regional centromere structure is found in most eukaryotes, including the closest related organisms with identified centromeres: the protist Plasmodium falciparum and the red alga C. merolae (21)(22)(23)(24). Both organisms have similarly sized centromeric DNA regions (∼2-4 kb) and also share low-GC content as a characteristic of their centromeres: ∼3% relative to the genome average GC of 21.8% in the case of P. falciparum (21,22), and 48.4% relative to the genome average GC of 55% for C. merolae (24). Interestingly, C. merolae, which is the only other alga with well-characterized centromeres and the closest relation to P. tricornutum of the organisms studied, has centromeres with a GC content that is low only compared with the genome average and not intrinsically, similar to P. tricornutum centromeres.…”

Section: Discussionmentioning

confidence: 99%

“…Centromeres of Schizosaccharomyces pombe and other yeast species feature an unconserved core of ATrich DNA sequence often surrounded by inverted repeats (17)(18)(19)(20). The centromeres of the protist Plasmodium have no apparent sequence similarity besides being 2-4-kb regions of extremely low-GC content (<3%) (21,22). Likewise, centromere regions of the red algal species Cyanidioschyzon merolae contain 2-3 kb of relatively low-GC content but manifest no other apparent pattern (23,24).…”

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Diatom centromeres suggest a mechanism for nuclear DNA acquisition

Diner

Noddings

Lian

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Centromeres are essential for cell division and growth in all eukaryotes, and knowledge of their sequence and structure guides the development of artificial chromosomes for functional cellular biology studies. Centromeric proteins are conserved among eukaryotes; however, centromeric DNA sequences are highly variable. We combined forward and reverse genetic approaches with chromatin immunoprecipitation to identify centromeres of the model diatom Phaeodactylum tricornutum. We observed 25 unique centromere sequences typically occurring once per chromosome, a finding that helps to resolve nuclear genome organization and indicates monocentric regional centromeres. Diatom centromere sequences contain low-GC content regions but lack repeats or other conserved sequence features. Native and foreign sequences with similar GC content to P. tricornutum centromeres can maintain episomes and recruit the diatom centromeric histone protein CENH3, suggesting nonnative sequences can also function as diatom centromeres. Thus, simple sequence requirements may enable DNA from foreign sources to persist in the nucleus as extrachromosomal episomes, revealing a potential mechanism for organellar and foreign DNA acquisition.diatom | Phaeodactylum tricornutum | episome | centromere | CENH3

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Diatom centromeres suggest a mechanism for nuclear DNA acquisition

Diner

Noddings

Lian

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…In this study we identify the centromeres of the chromosomes of T. gondii. Unlike Plasmodium (26,35), Toxoplasma centromeres show no nucleotide bias, and we did not identify obvious shared sequence motifs upon inspection of the primary sequence and bioinformatics motif searches. The centromeres are relatively small but have distinctive epigenetic features.…”

Section: Biochemical Confirmation Of Centromeres By Etoposide-mediatedmentioning

confidence: 99%

Toxoplasma gondii sequesters centromeres to a specific nuclear region throughout the cell cycle

Brooks

Francia

Gissot

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

161

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Members of the eukaryotic phylum Apicomplexa are the cause of important human diseases including malaria, toxoplasmosis, and cryptosporidiosis. These obligate intracellular parasites produce new invasive stages through a complex budding process. The budding cycle is remarkably flexible and can produce varied numbers of progeny to adapt to different host-cell niches. How this complex process is coordinated remains poorly understood. Using Toxoplasma gondii as a genetic model, we show that a key element to this coordination is the centrocone, a unique elaboration of the nuclear envelope that houses the mitotic spindle. Exploiting transgenic parasite lines expressing epitope-tagged centromeric H3 variant CenH3, we identify the centromeres of T. gondii chromosomes by hybridization of chromatin immunoprecipitations to genome-wide microarrays (ChIP-chip). We demonstrate that centromere attachment to the centrocone persists throughout the parasite cell cycle and that centromeres localize to a single apical region within the nucleus. Centromere sequestration provides a mechanism for the organization of the Toxoplasma nucleus and the maintenance of genome integrity.histone | mitosis

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“…In a previous study, we made a Plasmodium artificial chromosome (PAC) from the small centromere (1189 bp) and the short telomeric fragment (<250 bp) of the rodent malaria parasite Plasmodium berghei, which is widely used as a model for human malaria (Iwanaga et al 2010). This PAC is the second successful development of a functional eukaryotic artificial chromosome after the YAC.…”

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confidence: 99%

A high-coverage artificial chromosome library for the genome-wide screening of drug-resistance genes in malaria parasites

Iwanaga

Kaneko²

2012

Genome Res.

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The global spread of drug-resistant parasites is a serious problem for the treatment of malaria. Although identifying drugresistance genes is crucial for the efforts against resistant parasites, an effective approach has not yet been developed. Here, we report a robust method for identifying resistance genes from parasites by using a Plasmodium artificial chromosome (PAC). Large genomic DNA fragments (10-50 kb) from the drug-resistant rodent malaria parasite Plasmodium berghei were ligated into the PAC and directly introduced into the drug-sensitive (i.e., wild-type) parasite by electroporation, resulting in a PAC library that encompassed the whole genomic sequence of the parasite. Subsequently, the transformed parasites that acquired resistance were selected by screening with the drug, and the resistance gene in the PAC was successfully identified. Furthermore, the drug-resistance gene was identified from a PAC library that was made from the pyrimethamine-resistant parasite Plasmodium chabaudi, further demonstrating the utility of our method. This method will promote the identification of resistance genes and contribute to the global fight against drug-resistant parasites.

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Functional Identification of the Plasmodium Centromere and Generation of a Plasmodium Artificial Chromosome

Cited by 14 publications

References 0 publications

Diatom centromeres suggest a mechanism for nuclear DNA acquisition

Diatom centromeres suggest a mechanism for nuclear DNA acquisition

Toxoplasma gondii sequesters centromeres to a specific nuclear region throughout the cell cycle

A high-coverage artificial chromosome library for the genome-wide screening of drug-resistance genes in malaria parasites

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