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The Biology of Retrotransposition in Entamoeba histolytica

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Amebiasis

Abstract

Retrotransposons move into nonhomologous insertion sites within the genome via an RNA intermediate that is reverse transcribed into DNA. Those that lack long terminal repeats include long and short interspersed nuclear elements (LINEs and SINEs). LINEs are autonomous; partner SINEs retrotranspose using the LINE machinery. Entamoeba histolytica contains three classes of LINEs (EhLINE1, 2, 3) and SINEs (EhSINE1, 2, 3), which constitute approximately 11 % of the genome. EhLINE1 (4.8 kb) and EhSINE1 (550 bp) are the most abundant. They insert at AT-rich sites on all chromosomes, are not telomeric, and are close to protein-coding genes.

EhLINEs typically encode two open reading frames (ORFs). The N-terminal one-third of EhLINE1 contains ORF1, which has nucleic acid-binding properties. The ORF2 contains the reverse transcriptase (RT) domain and the endonuclease (EN) domain, which resembles type IIS restriction endonucleases. The purified EN domain protein could nick pBluescript DNA, and lacked strict sequence specificity. It displayed low K m, suggesting high affinity for DNA, and a low turnover number that could limit retrotransposition. Although EhLINE1 ORF1p is expressed in cultured E. histolytica cells, ORF2p is not detected. A cell line was obtained that expressed ORF2p by tetracycline induction and also contained an EhSINE1 copy marked with a GC-rich tag. In the presence of tetracycline, mobilization of the marked EhSINE1 was observed. Interestingly, mobilized EhSINE1 copies engaged in active sequence exchange during retrotransposition, probably brought about by multiple template jumping of RT, leading to rapid spread of sequence tag to the EhSINE1 population and generation of diversity.

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  1. School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Vijay Pal Yadav & Sudha Bhattacharya

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  1. Department of Parasitology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan

    Tomoyoshi Nozaki

  2. Jawaharlal Nehru University, New Delhi, India

    Alok Bhattacharya

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Yadav, V.P., Bhattacharya, S. (2015). The Biology of Retrotransposition in Entamoeba histolytica . In: Nozaki, T., Bhattacharya, A. (eds) Amebiasis. Springer, Tokyo. https://doi.org/10.1007/978-4-431-55200-0_10

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