Generation of Rodent Malaria Parasites with a High Mutation Rate by Destructing Proofreading Activity of DNA Polymerase δ
| Autor: | Satoru Kawai, Hiroyoshi Endo, Makoto Hirai, Shin-ichiro Kawazu, Mitsuru Furusawa, Toshihiro Horii, Shota Nakamura, Toshihiro Mita, Kazuyuki Tanabe, Hajime Honma, Nirianne Marie Q. Palacpac, Jun Ohashi, Hiroyuki Matsuoka, Hassan Hakimi, Teruo Yasunaga, Hajime Hisaeda |
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| Rok vydání: | 2014 |
| Předmět: |
Plasmodium
Mutation rate Sex Differentiation Plasmodium berghei DNA polymerase Mice 03 medical and health sciences 0302 clinical medicine Mutation Rate parasitic diseases Genetics medicine Gametocyte Animals Molecular Biology DNA Polymerase III 030304 developmental biology 0303 health sciences biology mutator Plasmodium falciparum DNA polymerase δ General Medicine Full Papers biology.organism_classification medicine.disease Virology Malaria 3. Good health genome sequencing biology.protein Proofreading Female 030217 neurology & neurosurgery |
| Zdroj: | DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes |
| ISSN: | 1756-1663 1340-2838 |
| DOI: | 10.1093/dnares/dsu009 |
| Popis: | Plasmodium falciparum malaria imposes a serious public health concern throughout the tropics. Although genetic tools are principally important to fully investigate malaria parasites, currently available forward and reverse tools are fairly limited. It is expected that parasites with a high mutation rate can readily acquire novel phenotypes/traits; however, they remain an untapped tool for malaria biology. Here, we generated a mutator malaria parasite (hereinafter called a 'malaria mutator'), using site-directed mutagenesis and gene transfection techniques. A mutator Plasmodium berghei line with a defective proofreading 3' → 5' exonuclease activity in DNA polymerase δ (referred to as PbMut) and a control P. berghei line with wild-type DNA polymerase δ (referred to as PbCtl) were maintained by weekly passage in ddY mice for 122 weeks. High-throughput genome sequencing analysis revealed that two PbMut lines had 175-178 mutations and a 86- to 90-fold higher mutation rate than that of a PbCtl line. PbMut, PbCtl, and their parent strain, PbWT, showed similar course of infection. Interestingly, PbMut lost the ability to form gametocytes during serial passages. We believe that the malaria mutator system could provide a novel and useful tool to investigate malaria biology. |
| Databáze: | OpenAIRE |
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