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The R2 mobile element of Rhynchosciara americana: Molecular, cytological and dynamic aspects

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Abstract

Ribosomal RNA genes are encoded by large units clustered (18S, 5S, and 28S) in the nucleolar organizer region in several organisms. Sometimes additional insertions are present in the coding region for the 28S rDNA. These insertions are specific non-long terminal repeat retrotransposons that have very restricted integration targets within the genome. The retrotransposon present in the genome of Rhynchosciara americana, RaR2, was isolated by the screening of a genomic library. Sequence analysis showed the presence of conserved regions, such as a reverse transcriptase domain and a zinc finger motif in the amino terminal region. The insertion site was highly conserved in R. americana and a phylogenetic analysis showed that this element belongs to the R2 clade. The chromosomal localization confirmed that the RaR2 mobile element was inserted into a specific site in the rDNA gene. The expression level of RaR2 in salivary glands during larval development was determined by quantitative RT-PCR, and the increase of relative expression in the 3P of the fourth instar larval could be related to intense gene activity characteristic of this stage. 5′-Truncated elements were identified in different DNA samples. Additionally, in three other Rhynchosciara species, the R2 element was present as a full-length element.

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Abbreviations

LINE:

long interspersed nuclear element

LTR:

long terminal repeat

mRNA:

messenger RNA

qPCR:

quantitative PCR

rDNA:

ribosomal DNA

rRNA:

ribosomal RNA

RT:

reverse transcriptase

SG:

salivary gland

SINE:

short interspersed nuclear element

UTR:

untranslated region

YL:

young larvae

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Acknowledgements

We thank Dr. Paolo M.A. Zanotto, Juliana Velasco and Fernando Lucas Melo for the sequencing support, and financial support and fellowship from FAPESP and CNPq.

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Authors and Affiliations

  1. Departamento de Biologia Celular e Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo, Avenida Professor Lineu Prestes, 1524, São Paulo, CEP 05508-900, SP, Brazil

    Paula Rezende-Teixeira, Fábio Siviero, Marina da Costa Rosa & Glaucia Maria Machado-Santelli

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  1. Paula Rezende-Teixeira
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  2. Fábio Siviero
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  3. Marina da Costa Rosa
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  4. Glaucia Maria Machado-Santelli
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Correspondence to Paula Rezende-Teixeira.

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Responsible Editor: Herbert Macgregor

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Fig. S1

Nucleotide sequence and conceptual translation of the consensus sequence of the RaR2 non-LTR retrotransposon of Rhynchosciara americana. The insertion site is shown in 28 S rDNA and some conserved motifs such as the CCHH amino acid motif with double underline and the reverse transcriptase domain in italic bold. The main reverse transcriptase domains are underlined and numbered in the sequence. In the carboxyl-terminal region, the CCHC motif is dotted and the KPDLV conserved domain is in gray (PDF 26 kb)

Fig. S2

Alignment of the amino acid sequences of the R2 retrotransposon ORFs of Rhynchosciara americana (R. americana), Bradysia coprophila (B. coprophila), and Bombyx mori (Burke et al. 1987, 1993): Gray and black boxes indicate identical and conserved replacement amino acids (PDF 21 kb)

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Rezende-Teixeira, P., Siviero, F., da Costa Rosa, M. et al. The R2 mobile element of Rhynchosciara americana: Molecular, cytological and dynamic aspects. Chromosome Res 17, 455–467 (2009). https://doi.org/10.1007/s10577-009-9038-x

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