962 resultados para 5S RDNA


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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Samples from seven different locations of the genus Pimelodella were genetically examined, two caves (exclusively subterranean, upper Tocantins River and Sao Francisco River) and five epigean (from upper Parana River basin). Cytogenetic analyses revealed the same diploid number (2n=46) for all species besides similarities in both number and location of nucleolar organizer regions and C bands. FISH with 5S rDNA probes and CMA(3) staining indicated significant differences among the studied species. Application of PCR-RFLP in ATPase 6 and 8 mitochondrial genes allowed building a minimum evolution phenogram identifying the close evolutionary relationship among groups. Both chromosomal and molecular data were useful to infer the relationships among studied Pimelodella species.

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Defined model systems consisting of physiologically spaced arrays of H3/H4 tetramer⋅5S rDNA complexes have been assembled in vitro from pure components. Analytical hydrodynamic and electrophoretic studies have revealed that the structural features of H3/H4 tetramer arrays closely resemble those of naked DNA. The reptation in agarose gels of H3/H4 tetramer arrays is essentially indistinguishable from naked DNA, the gel-free mobility of H3/H4 tetramer arrays relative to naked DNA is reduced by only 6% compared with 20% for nucleosomal arrays, and H3/H4 tetramer arrays are incapable of folding under ionic conditions where nucleosomal arrays are extensively folded. We further show that the cognate binding sites for transcription factor TFIIIA are significantly more accessible when the rDNA is complexed with H3/H4 tetramers than with histone octamers. These results suggest that the processes of DNA replication and transcription have evolved to exploit the unique structural properties of H3/H4 tetramer arrays.

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A base-pair resolution method for determining nucleosome position in vitro has been developed to com- plement existing, less accurate methods. Cysteaminyl EDTA was tethered to a recombinant histone octamer via a mutant histone H4 with serine 47 replaced by cysteine. When assembled into nucleosome core particles, the DNA could be cut site specifically by hydroxyl radical-catalyzed chain scission by using the Fenton reaction. Strand cleavage occurs mainly at a single nucleotide close to the dyad axis of the core particle, and assignment of this location via the symmetry of the nucleosome allows base-pair resolution mapping of the histone octamer position on the DNA. The positions of the histone octamer and H3H4 tetramer were mapped on a 146-bp Lytechinus variegatus 5S rRNA sequence and a twofold-symmetric derivative. The weakness of translational determinants of nucleosome positioning relative to the overall affinity of the histone proteins for this DNA is clearly demonstrated. The predominant location of both histone octamer and H3H4 tetramer assembled on the 5S rDNA is off center. Shifting the nucleosome core particle position along DNA within a conserved rotational phase could be induced under physiologically relevant conditions. Since nucleosome shifting has important consequences for chromatin structure and gene regulation, an approach to the thermodynamic characterization of this movement is proposed. This mapping method is potentially adaptable for determining nucleosome position in chromatin in vivo.

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The Acanthuridae family is a representative group from the marine fish that plays a key role in ecological dynamics of coral reefs. Three species are common along coastal reefs of Western Atlantic: Acanthurus coeruleus, Acanthurus bahianus and Acanthurus chirurgus. In the present study, cytogenetic data are presented for these three species Acanthurus based on classical cytogenetic methods and mapping of repetitive sequences such as ribosomal 18S and 5S rDNA and telomeric repeats to improve their karyotype evolutionary analyses. The cytogenetic pattern of these species indicated sequential steps of chromosomal rearrangements dating back 19 to 5 millions of years ago (M.a.) that accounted for their interspecific differences. A. coeruleus (2n=48; 2sm+4st+42a), A. bahianus (2n=36; 12m+2sm+4st+18a) and A. chirurgus (2n=34; 12m+2sm+4st+16a) share an older set of three chromosomal pairs that were originated through pericentric inversions. A set of six large metacentric pairs formed by Robertsonian (Rb) translocations found in A. bahianus and A. chirurgus and a putative in tandem fusion found in A. chirurgus are more recent events. The lack of interstitial telomeric sequences (ITS) in spite of several centric fusions in A. bahianus and A. chirurgus might be related to the long period of time after their occurrence (estimated in 5 M.a.). Furthermore, the homeologies among the chromosome pairs bearing ribosomal genes, in addition to other structural features, highlight large conserved chromosomal regions in the three species. Our findings indicate that macrostructural changes occurred during the cladogenesis of these species were not followed by conspicuous microstructural rearrangements in the karyotypes.