Reverse transcriptase-related proteins in telomeres and in certain chromosomal loci of Rhynchosciara (Diptera: Sciaridae).
The localization of reverse transcriptase-related proteins in polytene chromosomes of dipterans was investigated using previously characterized antibodies to a recombinant polypeptide containing conserved motifs of insect reverse transcriptases. The immunoreactions were carried out with polytene chromosome squashes of eight sciarids, one chironomid and three Drosophila species. Telomeric staining was regularly observed on chromosomes of the sciarid Rhynchosciara americana under normal growth conditions. Five of eight chromosomal tips were labelled except for the heterochromatic ends that are occasionally found associated forming a chromocentre in the salivary gland. Reverse transcriptase-related proteins were detected at chromosomal tips of young larvae and remained bound to the telomeres throughout larval development. As in salivary gland chromosomes, five non-telocentric ends of the chromosomes from Malpighian tubules of R. americana appeared clearly stained with anti-reverse transcriptase. The occurrence of telomeric reverse transcriptase in R. americana correlates with the presence of RNA in addition to an unusual enrichment with homopolymeric dA/dT DNA associated with the telomeric heterochromatin. The antibodies also reacted with a few interstitial sites in chromosomes of four Rhynchosciara species, one band overlapping the histone gene locus of three species in the americana -like group. The results provide evidence for a reverse transcriptase-related protein as a constitutive component in telomeres of R. americana and also in certain interstitial loci of Rhynchosciara species in which RNA was immunologically detected in the form of RNA:DNA hybrids.
Gorab
Eduardo
E
Departamento de Biologia, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, Cidade Universitária, Brazil CEP 05508-900. egorab@usp.br
D000818:Animals; D002843:Chromatin / Q000378:metabolism; D002875:Chromosomes / Q000648:ultrastructure; D004247:DNA / Q000737:chemistry; D004175:Diptera; D004330:Drosophila; D015151:Immunoblotting; D017403:In Situ Hybridization; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D012469:Salivary Glands / Q000378:metabolism; D016615:Telomere / Q000378:metabolism* / Q000648:ultrastructure
D002843:Chromatin; D004247:DNA; D012194:RNA-Directed DNA Polymerase