5S Clusters Research Articles

Cytogenetic characterization and mapping of the repetitive DNAs in Cycloramphus bolitoglossus (Werner, 1897): More clues for the chromosome evolution in the genus Cycloramphus (Anura, Cycloramphidae).

Cycloramphus bolitoglossus (Werner, 1897) is a rare species with a low population density in the Serra do Mar region of Paraná and Santa Catarina, in southern Brazil. Currently, it has been assigned to the Near Threatened (NT) category in the Brazilian List of Endangered Animal Species. Here, we described the karyotype of this species for the first time and investigated the patterns of some repetitive DNA classes in the chromosomes using molecular cytogenetic approaches. We isolated, sequenced and mapped the 5S rDNA and the satellite DNA PcP190 of C. bolitoglossus, as well as mapped the telomeric sequences and seven microsatellites motifies [(GA)15, (CA)15, (GACA)4, (GATA)8, (CAG)10, (CGC)10, and (GAA)]10. Cycloramphus bolitoglossus has 2n = 26 chromosomes and a fundamental number (FN) equal to 52, with a highly conserved karyotype compared to other genus members. Comparative cytogenetic under the phylogenetic context of genus allowed evolutionary interpretations of the morphological changes in the homologs of pairs 1, 3, and 6 along with the evolutionary history of Cycloramphus. Two subtypes of 5S rDNA type II were isolated in C. bolitoglossus genome, and several comparative analysis suggests mixed effects of concerted and birth-and-death evolution acting in this repetitive DNA. The 5S rDNA II subtype "a" and "b" was mapped on chromosome 1. However, their different position along chromosome 1 provide an excellent chromosome marker for future studies. PcP190 satellite DNA, already reported for species of the families Hylidae, Hylodidae, Leptodactylidae, and Odontophrynidae, is scattered throughout the C. bolitoglossus genome, and even non-heterochromatic regions showed hybridization signals using the PcP190 probe. Molecular analysis suggests that PcP190 satellite DNA exhibit a high-level of homogenization of this sequence in the genome of C. bolitoglossus. The PcP190 satDNA from C. bolitoglossus represents a novel sequence group, compared to other anurans, based on its hypervariable region. Overall, the present data on repetitive DNA sequences showed pseudogenization evidence and corroborated the hypothesis of the emergence of satDNA from rDNA 5S clusters. These two arguments that reinforced the importance of the birth-and-death evolutionary model to explain 5S rDNA patterns found in anuran genomes.

Evolutionary Dynamics of Copy Number and Meiotic Recombination in Murine 5S rDNA: Possible Involvement of Natural Selection.

We investigated evolutionary trends of the 5S ribosomal RNA gene in the house mouse, Mus musculus. First, we assessed the 5S cluster and copy numbers in eight laboratory strains by pulsed-field gel electrophoresis. The copy numbers in seven lines were estimated to be around 130-170 copies per cluster, with 63 copies in the remaining strain, implying that the copy number can change drastically and has been maintained under certain evolutionary constraints at ~ 140 copies. Second, we addressed the frequency of meiotic recombination mediated by the 5S cluster by performing a mating experiment with laboratory strains, and found that the 5S cluster did not accelerate recombination events. Third, we surveyed recombination events of the 5S-containing chromosome region in wild mice from the Japanese Islands, where the two subspecies lineages, M. m. castaneus and M. m. musculus, are historically mingled, and found that the influence of the 5S cluster on meiotic recombination was limited. Finally, we examined the nucleotide diversity of six genes in the neighboring regions of the 5S cluster and found reduced genetic diversity in the regions on both sides of the cluster, suggesting the involvement of either positive or background selection in the population-level sequence similarity of the 5S clusters. Therefore, the mouse 5S genes are considered to be evolving toward sequence similarity within a given cluster by certain intrachromosomal mechanisms and toward sharing of a specific 5S cluster within a population by certain selective processes.

Cytogenetic Mapping of H1 Histone and Ribosomal RNA Genes in Hybrids between Catfish Species Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum

A physical chromosome mapping of the H1 histone and 5S and 18S ribosomal RNA (rRNA) genes was performed in interspecific hybrids of Pseudoplatystoma corruscans and P. reticulatum. The results showed that 5S rRNA clusters were located in the terminal region of 2 chromosomes. H1 histone and 18S ribosomal genes were co-localized in the terminal portion of 2 chromosomes (distinct from the chromosomes bearing 5S clusters). These results represent the first report of association between H1 histone and 18S genes in fish genomes. The chromosome clustering of ribosomal and histone genes was already reported for different organisms and suggests a possible selective pressure for the maintenance of this association.

5S rDNA chromosomal mapping and COI sequence analysis reveal differentiation among distinct populations of a characid fish Serrapinnus notomelas

Cheirodontinae comprises a small-sized fish group with a wide geographical distribution throughout Central and South America, mainly in Brazilian Basins. The species Serrapinnus notomelas is widely distributed in the Upper Parana River Basin. In this work, chromosomal mapping of 5S ribosomal RNA (rRNA) gene was performed trough fluorescence in situ hybridization (FISH) in speciemens from three distinct localities belonging to two basins in the Sao Paulo State. All populations presented 5S clusters in two chromosome pairs. One additional pair was detected in samples from the Rio Paraitinguinha (Tiete Basin), and two additional pairs were detected in the population from Corrego Campo Novo (Tiete Basin). Analyses with partial sequences of COI were performed to verify the interelationship among the studied specimens, revealing Corrego Campo Novo population as a sister-group to the clade formed by the two other populations. The samples from Rio Paraitinguinha and Recanto dos Cambaras presented two distinct haplotypes each, while five haplotypes were observed in the Corrego Campo Novo population, sugesting that this could be older than the other populations. None of the nine haplotypes were shared among the three populations. The similarities and differences observed among the three populations using cytogenetic data and COI analysis are not related to geographic distances that separate the samples, suggesting that the origin of the Rio Paraitinguinha and Recanto dos Cambaras populations may be related to faunal exchanges. Additionally, the present data suggest that the analyzed populations of S. notomelas may be on independent evolutionary trajectories but in a very initial diversification stage. Moreover, the use of integrative information, such as molecular and chromosomal data in the analysis of population divergence and evolutionary trajectories in freshwater fishes is reinforced.

Cytogenetic and molecular analysis of the pufferfish Tetraodon fluviatilis (Osteichthyes).

In view of their compact genome, pufferfish (Tetraodontiformes) have been proposed as model animal for the study of the vertebrate genome. Despite such interest, cytogenetic information about puffers is still scanty. To fill this gap, a cytogenetic analysis of T. fluviatilis has been performed using both classical and molecular techniques. C-banding, followed by DAPI staining, evidenced that in T. fluviatilis, like all other puffer species so far examined, heterochromatin is essentially AT-rich and it is located at centromeres, whereas staining with CMA3, silver staining and FISH with a 28S ribosomal RNA gene DNA probe showed 2-4 nucleolar organizing regions (NORs) located in heterochromatic regions in the considered puffer species. FISH with the 5S probe put in evidence both in T. fluviatilis and in T. nigroviridis only a 5S cluster per haploid genome that is physically unlinked with the major ribosomal RNA genes including the 28S rRNA genes. Hybridization with the (TTAGGG)n probe showed in all the puffers brightly fluorescent signals uniform both in size and intensity at the end of all the chromosomes. Finally, mariner-like elements (MLEs) have been identified in T. fluviatilis and they have located into the NOR-associated heterochromatin.

An approach to study the evolution of theDrosophila 5S ribosomal genes using P-element transformation

The P-element-mediated gene transfer system was used to introduce Drosophila teissieri 5S genes into the Drosophila melanogaster genome. Eight transformed D. melanogaster strains that carry D. teissieri 5S mini-clusters consisting of 9-21 adjacent 5S units were characterized. No genetic exchanges between D. melanogaster and D. teissieri 5S clusters were detected over a 2-year survey of the eight strains. The occurrence of small rearrangements within the D. melanogaster 5S cluster was demonstrated in one of the transformed strains.

Bipartite structure of the 5S ribosomal gene family in a Drosophila melanogaster strain, and its evolutionary implications.

Knowledge of multigenic family organization should provide insight into their mode of evolution. Accordingly, we characterized the 5S ribosomal gene family in the Drosophila melanogaster strain ry506. The 5S genes in this strain display a striking HindIII restriction difference compared to the "standard" D. melanogaster 5S genes. The sequence of three ry506 5S genes was determined. We show that the HindIII restriction site heterogeneity within the ry506 5S family most probably results from the same point mutation, suggesting that a single 5S variant was propagated into the 5S cluster of this strain. Furthermore, we demonstrate that the structural organization of the 5S genes in ry506 is a bipartite structure, i.e., that about 40% of the 5S genes constitute a HindIII+/HindIII- mixed cluster, while those remaining constitute an homogeneous HindIII- cluster. The events which might lead to such an heterogeneous pattern are discussed from an evolutionary point of view.

Sites of the 5S ribosomal genes in Drosophila. I. The multiple clusters in the virilis group.

Drosophila melanogaster 125I-5S RNA was annealed to salivary gland preparations of 6 species in the virilis group of Drosophila. Two patterns of annealing were found. D. virilis, D. montana and D. borealis showed three 5S gene clusters on chromosome 5; Sd-f and Wc-j were strongly labeled, but Xa-e was weakly labeled. D. montana and D. borealis have a greater percentage of their total 5S cistroms at S d-f than does D. virilis. D. americana americana, D. americana texana and D. novamexicana showed 2 sites labeled; no label was seen at Sd-f while Wc-j was weakly labeled and Xa-e was strongly labeled, the reciprocal of the previous pattern in the W-X region. Hybrids between D. a americana and D. virilis showed no difference in chromosome banding at the sites of the 5S clusters despite their pattern difference. D. a texana X D. virilis, on the other hand, did show a difference in staining the Xa-e region. These patterns fall squarely into the biosystematic groupings deduced by many previous workers.