Abstract

BackgroundFor anurans, knowledge of 5S rDNA is scarce. For Engystomops species, chromosomal homeologies are difficult to recognize due to the high level of inter- and intraspecific cytogenetic variation. In an attempt to better compare the karyotypes of the Amazonian species Engystomops freibergi and Engystomops petersi, and to extend the knowledge of 5S rDNA organization in anurans, the 5S rDNA sequences of Amazonian Engystomops species were isolated, characterized, and mapped.ResultsTwo types of 5S rDNA, which were readily differentiated by their NTS (non-transcribed spacer) sizes and compositions, were isolated from specimens of E. freibergi from Brazil and E. petersi from two Ecuadorian localities (Puyo and Yasuní). In the E. freibergi karyotypes, the entire type I 5S rDNA repeating unit hybridized to the pericentromeric region of 3p, whereas the entire type II 5S rDNA repeating unit mapped to the distal region of 6q, suggesting a differential localization of these sequences. The type I NTS probe clearly detected the 3p pericentromeric region in the karyotypes of E. freibergi and E. petersi from Puyo and the 5p pericentromeric region in the karyotype of E. petersi from Yasuní, but no distal or interstitial signals were observed. Interestingly, this probe also detected many centromeric regions in the three karyotypes, suggesting the presence of a satellite DNA family derived from 5S rDNA. The type II NTS probe detected only distal 6q regions in the three karyotypes, corroborating the differential distribution of the two types of 5S rDNA.ConclusionsBecause the 5S rDNA types found in Engystomops are related to those of Physalaemus with respect to their nucleotide sequences and chromosomal locations, their origin likely preceded the evolutionary divergence of these genera. In addition, our data indicated homeology between Chromosome 5 in E. petersi from Yasuní and Chromosomes 3 in E. freibergi and E. petersi from Puyo. In addition, the chromosomal location of the type II 5S rDNA corroborates the hypothesis that the Chromosomes 6 of E. petersi and E. freibergi are homeologous despite the great differences observed between the karyotypes of the Yasuní specimens and the others.

Highlights

  • Molecular Organization of the 5S ribosomal DNA (rDNA) in Engystomops This study verified the occurrence of two types of 5S rDNA in the genomes of the Amazonian species of Engystomops, a feature widely documented in many vertebrates, including fish [10,11,36,37,38,39,40,41,42], the anurans Xenopus laevis and Xenopus borealis [6,16,21,22,23,24,43,44] chickens [45,46,47], and mammals [12,48,49]

  • To the findings reported in these studies, the two types of 5S rDNA sequences found in E. freibergi and E. petersi varied slightly in their corresponding coding regions, with the main difference between them found in the non-transcribed spacer (NTS) region, which varied in length (84 bp for the type I 5S rDNA and approximately 650 bp for the type II 5S rDNA) and nucleotide composition

  • These two types of 5S rDNA do not, appear to be related to the dual system observed in Xenopus [6,13,16,20,21,22,24] because we found no similarities between the NTSs of the oocyte- or somatictype sequences to those in the present study

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Summary

Introduction

Knowledge of 5S rDNA is scarce. For Engystomops species, chromosomal homeologies are difficult to recognize due to the high level of inter- and intraspecific cytogenetic variation. Within the conserved 120bp sequence lies an internal control region (ICR) consisting of 3 characteristic regions: the A box, an intermediate element (IE), and the C box [5] These regions act as promoters for transcription; the A box is a general binding sequence for RNA polymerase III, and the intermediate element and the C box are interaction sites for the transcription factor TFIIIA [5]. Another characteristic typically found in the presumed-functional 5S genes is a poly-T terminator region, as initially reported by Korn and Brown [6]

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