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Replication pattern of human repeated DNA sequences.

Either aphidicolin- or thymidine-synchronized human HL-60 cells were used to study the replication pattern of a family of human repetitive DNA sequences, the Eco RI 340 bp family (alpha RI-DNA), and of the ladders of fragments generated in total human DNA after digestion with XbaI and HaeIII (alpha satellite sequences). DNAs replicated in early, middle-early, middle-late and late S periods were labelled with BUdR or with [3H]thymidine. The efficiency of the cell synchronization procedure was confirmed by the transition from a high-GC to a high-AT average base composition of the DNA synthesized going from early to late S periods. By hybridizing EcoRI 340 bp repetitive fragments to BUdR-DNAs it was found that this family of sequences is replicated throughout the entire S period. Comparing fluorograph densitometric scans of [3H]DNAs to the scans of ethidium bromide patterns of total HL-60 DNA digested with XbaI and HaeIII, it was observed that DNA synthesized in different S periods is characterized by approximately the same ladder of fragments, while the intensity of each band may vary through the S phase; in particular, the XbaI 2.4 kb fragment becomes undetectable in late S.

Base Sequence↗

First- or second-order transition in the melting of repeat sequence DNA.

Both theoretical analysis and observation of the continuity of the melted fraction of base pairs indicate that the melting transition in DNA is second order. Analysis of the salt dependence of the transition by polyelectrolyte limiting laws, however, has first-order dynamics imbedded in the analysis. This paper proposes that the observation taken to be a latent heat of melting in the limiting law analysis could instead be a specific heat anomaly associated with a second-order transition. The limiting laws can be reconstructed based on a second-order transition with a specific heat anomaly. The T2M dependence of this excess heat is also consistent with its being a specific heat anomaly of a system displaying classical critical behavior. Classical critical behavior indicates that theoretical mean field approaches such as MSPA should be particularly appropriate to helix melting studies.

DNA↗

Species-specific evolution of repeated DNA sequences in great apes.

DNA sequencing reveals that the genomes of the human, gorilla and chimpanzee share more than 98% homology. Comparative chromosome painting and gene mapping have demonstrated that only a few rearrangements of a putative ancestral mammalian genome occurred during great ape and human evolution. However, interspecies representational difference analysis (RDA) of the gorilla between human and gorilla revealed gorilla-specific DNA sequences. Cloning and sequencing of gorilla-specific DNA sequences indicate that there are repetitive elements. Gorilla-specific DNA sequences were mapped by fluorescence in-situ hybridization (FISH) to the subcentromeric/centromeric regions of three pairs of gorilla submetacentric chromosomes. These sequences could represent either ancient sequences that got lost in other species, such as human and orang-utan, or, more likely, recent sequences which evolved or originated specifically in the gorilla genome.

Animals↗

Repeated DNA sequences in the microbat species Miniopterus schreibersi (Vespertilionidae; Chiroptera).

Repetitive DNA sequences represent a substantial component of eukaryotic genomes. These sequences have been described and characterized in many mammalian species. However, little information about repetitive DNA sequences is available in bat species. Here we describe an EcoRI family of repetitive DNA sequences present in the species Miniopterus schreibersi. These repetitive sequences are 57.85%, A-T rich, organized in tandem, and with a monomer unit length of 904 bp. Methylation analysis using the isoesquizomer pair MspI and HpaII indicates that the cytosines present in the sequences CCGG are partially methylated. Furthermore, Southern blot analysis demonstrated that these DNA sequences are absent in the genomes of four related microbat species and suggest that it could be specific to the M. schreibersi genome.

Animals↗

Highly repeated DNA sequences in three species of the genus Pteropus (Megachiroptera, Mammalia).

Bat genomes are characterised by an A-T richness and by a small C-value compared with other mammalian groups. It has been suggested that the small C-value is mainly due to the lack of repetitive DNA sequences. However, little information about repetitive DNA sequences in this mammalian group is available at the molecular level. Here we describe a PstI family of repetitive DNA sequences present in three species of the genus Pteropus. These repetitive sequences are 54.97% G-C rich, organised in tandem and with a unit length of 744 bp. Methylation analysis indicates that some of the CCGG target sites present in these repetitive DNA sequences have methylated cytosines and that there are small differences in the methylation pattern between species. Several features of this family of repetitive sequences suggest that they evolved by concerted evolution.

Animals↗

Genetic organization of a repeated DNA sequence family in the rice blast fungus.

The fungal rice pathogen Magnaporthe grisea contains repetitive DNA sequences called MGR. We have used a DNA probe, MGR586, derived from these sequences and crosses between rice-pathogenic and non-rice-pathogenic laboratory strains of M. grisea to rapidly map genes in this organism. The rice-pathogenic strain contained 57 EcoRI restriction fragments that hybridize to the MGR586 probe; the other five non-rice-pathogenic parent strains contained a single MGR586 sequence. Genetic analysis of MGR segregation detected eight linkage groups and allowed the mapping of three pigmentation genes (Alb1, Rsy1, and Buf1), the mating type locus (Mat1), the nucleolar organizer (Rdn1), the Smol gene, and two restriction fragment length polymorphisms linked to Smol. Our results indicate that the MGR586 loci are randomly distributed about the M. grisea genome and permit the construction of a well-marked linkage map useful for future studies on genome organization and genetic analysis in M. grisea.

Journal Article↗

Species-specific homogeneity of the primate Alu family of repeated DNA sequences.

We have determined the base sequence of several cloned Alu family members from the DNAs of a new world monkey (owl monkey) and a prosimian (galago). The three owl monkey Alu family members reported here belong to a single 300 base pair consensus sequence which closely resembles the human Alu family consensus. The galago Alu family members can best be represented as belonging to either of two related but distinct consensus sequences. One of the two galago Alu family subgroups (Type I) more accurately resembles the human consensus sequence than does the other subgroup (Type II). In this work we compare base sequences of human and galago Type I Alu family members. There are several examples of species-specific differences between the human and Type I galago sequences indicating that the Alu family members are effectively homogenized within a species.

Animals↗