Molecular characterization of a retrotransposon in the Rhynchosciara americana genome and its association with telomere.
Non-LTR retrotransposons, also known as long interspersed nuclear elements (LINEs), are transposable elements that encode a reverse transcriptase and insert into genomic locations via RNA intermediates. The sequence analysis of a cDNA library constructed from mRNA of the salivary glands of R. americana showed the presence of putative class I elements. The cDNA clone with homology to a reverse transcriptase was the starting point for the present study. Genomic phage was isolated and sequenced and the molecular structure of the element was characterized as being a non-LTR retrotransposable element. Southern blot analysis indicated that this transposable element is represented by repeat sequences in the genome of R. americana. Chromosome tips were consistently positive when this element was used as probe in in-situ hybridization. Real-time RT-PCR showed that this retrotransposon is transcribed at different periods of larval development. Most interesting, the silencing of this retrotransposon in R. americana by RNA interference resulted in reduced transcript levels and in accelerated larval development.
Departamento de Biologia Celular e do Desenvolvimento, Avenida Professor Lineu Prestes, 1524 Y ICBI Y sala 307, Universidade de São Paulo, São Paulo, Brazil. paularez@usp.br
Paula
P
Rezende-Teixeira
Fábio
F
Siviero
Amanda S
AS
Brandão
Roberto Vicente
RV
Santelli
Gláucia M
GM
Machado-Santelli
D000818:Animals; D004175:Diptera / Q000235:genetics* / Q000254:growth & development; D020868:Gene Silencing; D017403:In Situ Hybridization; D020084:Long Interspersed Nucleotide Elements / Q000235:genetics*; D008957:Models, Genetic; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012469:Salivary Glands; D017422:Sequence Analysis, DNA; D016615:Telomere / Q000502:physiology*