Bayesian inference phylogenetic tree based on Hsp90 protein sequences from Chordata.

<p>Species in which multiple <i>hsp90</i> genes have been detected are in dark blue. Filled diamonds denote putative species-specific duplication events, predicted by this study. Filled square denotes the duplication event resulting in the cognate and inducible isoforms of vertebrates. Numbers at nodes represent Posterior Probability (PP) values. Scale bar: substitutions/site.</p

Bayesian inference phylogenetic tree based on Hsp90 protein sequences from Mollusca.

<p>Species in which multiple <i>hsp90</i> genes have been detected are in dark blue. Filled diamonds denote putative species-specific duplication events, predicted by this study. Numbers at nodes represent Posterior Probability (PP) values. Scale bar: substitutions/site.</p

Assessment and Reconstruction of Novel HSP90 Genes: Duplications, Gains and Losses in Fungal and Animal Lineages

<div><p>Hsp90s, members of the Heat Shock Protein class, protect the structure and function of proteins and play a significant task in cellular homeostasis and signal transduction. In order to determine the number of <i>hsp90</i> gene copies and encoded proteins in fungal and animal lineages and through that key duplication events that this family has undergone, we collected and evaluated Hsp90 protein sequences and corresponding Expressed Sequence Tags and analyzed available genomes from various taxa. We provide evidence for duplication events affecting either single species or wider taxonomic groups. With regard to Fungi, duplicated genes have been detected in several lineages. In invertebrates, we demonstrate key duplication events in certain clades of Arthropoda and Mollusca, and a possible gene loss event in a hymenopteran family. Finally, we infer that the duplication event responsible for the two (a and b) isoforms in vertebrates occurred probably shortly after the split of Hyperoartia and Gnathostomata.</p></div

Bayesian inference phylogenetic tree based on Hsp90 protein sequences from Fungi.

<p>Species in which multiple <i>hsp90</i> genes have been detected are in dark blue. Filled diamonds denote putative species-specific duplication events, predicted by this study. Stars represent whole-genome duplications reported by previous studies. Numbers at nodes represent Posterior Probability (PP) values. Scale bar: substitutions/site.</p

Bayesian inference phylogenetic tree based on Hsp90 protein sequences from Arthropoda.

<p>Species in which multiple <i>hsp90</i> genes have been detected are in dark blue. Filled diamonds denote putative species-specific duplication events predicted by this study. Filled squares denote duplication events in the common ancestor of a wide taxonomic group (e.g. Pleocyemata), predicted by this study. Filled circle shows the gain of type 2 isoform in Hymenoptera; empty-strikethrough circle shows loss of type 1 isoform in Formicidae. Numbers at nodes represent Posterior Probability (PP) values. Scale bar: substitutions/site.</p

Species distribution of BoR300 repeats.

<p>HaeIII-digested genomic DNA of representative species of the Diptera family and the genus Bactrocera was analyzed by Southern hybridization using as probe the biotinylated monomer (298 bp) of the repeat. (<b>A</b>) Southern blot analysis of digested genomic DNA from the following dipteran species: <i>B. oleae</i> (lane 1), <i>C. capitata</i> (lane 2), <i>D. melanogaster</i> (lane 3) and <i>An. gambiae</i> (lane 4). <b>B</b>) Southern blot analysis of digested genomic DNA from the following Bactrocera species: <i>B. oleae</i> (lane 1), <i>B. correcta</i> (lane 3), <i>B. cucurbitae</i> (lane 4), and <i>B. dorsalis</i> (lane 5). In Lane 2 is the cloned monomer of the repeat. M represents the molecular marker (SM0331, Fermentas).</p

qPCR analysis data of the relative estimation of BoR300 repeats using a reference standard curve.

a<p>Initial template concentration of the <i>B</i>. <i>oleae</i> genomic DNA (pg) used at the qPCR reactions.</p>b<p>Mean copy number of BoR300 repeats which was estimated for the initial template concentration of the genomic DNA (a) based on the standard curve.</p>c<p><i>B. oleae</i> haploid genome size: 0.352 pg.</p>d<p>Standard Error (SE) for the triplicate measurements (n = 3).</p

FISH with the BoR300 probe on mitotic and polytene nuclei of <i>Bactrocera oleae.</i>

<p>Chromosomes were counterstained with DAPI (blue). Female (a) and male (b) metaphase showing strong hybridization signals (red) on the centromeres of chromosomes 4 and 5. Polytene complement (c) showing strong hybridization signals (red) on the centromeric heterochromatic blocks (C) of chromosomes III and IV (arrows). Bar  = 3 µm (a, b), 20 µm (c).</p

CENP-B like motif.

<p>Comparison of the CENP-B like motif (237–252 bp) found in BoR300 with the degenerate motif considered to bind the CENP-B protein.</p

Nucleotide sequence of the monomer BoR300.

<p>The arrows indicate the outfacing primer pair: the reverse (BoR300-R) and the forward (BoR300-F) primer respectively. The restriction sites of the restriction endonucleases HaeIII and TaqI are also highlighted.</p