Mitochondrial DNA of the extinct quagga: Relatedness and extent of postmortem change

Sequences are reported for portions of two mitochondrial genes from a domestic horse and a plains zebra and compared to those published for a quagga and a mountain zebra. The extinct quagga and plains zebra sequences are identical at all silent sites, whereas the horse sequence differs from both of them by 11 silent substitutions. Postmortem changes in quagga DNA may account for the two coding substitutions between the quagga and plains zebra sequences. The hypothesis that the closest relative of the quagga is the domestic horse receives no support from these data. From the extent of sequence divergence between horse and zebra mitochondrial DNAs (mtDNAs), as well as from information about the fossil record, we estimate that the mean rate of mtDNA divergence in Equus is similar to that in other mammals, i.e., roughly 2% per million years.     

Centrosomes, and not nuclei, initiate pole cell formation in Drosophila embryos

An injection of aphidicolin into early Drosophila embryos inhibits DNA synthesis and nuclear division, while centrosome replication and many other aspects of the mitotic cycle continue. If aphidicolin is injected at nuclear cycle 7-8, the normal migration of nuclei to the embryo cortex is completely inhibited. In most of these embryos, however, centrosomes continue to migrate in a coordinated manner to the cortex, where they reorganize tubulin, actin, and the overlying plasma membrane. Remarkably, the centrosomes that migrate to the posterior pole of such embryos initiate pole cell formation in the absence of nuclei. These observations demonstrate that centrosomes alone are able to direct a major reorganization of the cortical cytoskeleton when they arrive at the surface of the embryo. They also suggest that the coordinated movement of nuclei to the embryo cortex is mediated by forces acting on the centrosome rather than on the nucleus itself.     

Towards a physical map of the Drosophila melanogaster genome: mapping of cosmid clones within defined genomic divisions.

A physical map of the D. melanogaster genome is being constructed, in the form of overlapping cosmid clones that are assigned to specific polytene chromosome sites. A master library of ca. 20,000 cosmids is screened with probes that correspond to numbered chromosomal divisions (ca. 1% of the genome); these probes are prepared by microdissection and PCR-amplification of individual chromosomes. The 120 to 250 cosmids selected by each probe are fingerprinted by Hinfl digestion and gel electrophoresis, and overlaps are detected by computer analysis of the fingerprints, permitting us to assemble sets of contiguous clones (contigs). Selected cosmids, both from contigs and unattached, are then localized by in situ hybridization to polytene chromosomes. Crosshybridization analysis using end probes links some contigs, and hybridization to previously cloned genes relates the physical to the genetic map. This approach has been used to construct a physical map of the 3.8 megabase DNA in the three distal divisions of the x chromosome. The map is represented by 181 canonical cosmids, of which 108 clones in contigs and 32 unattached clones have been mapped individually by in situ hybridization to chromosomes. Our current database of in situ hybridization results also includes the beginning of a physical map for the rest of the genome: 162 cosmids have been assigned by in situ hybridization to 129 chromosomal subdivisions elsewhere in the genome, representing 5 to 6 megabases of additional mapped DNA.     

Increased frequency of 6-thioguanine-resistant peripheral blood lymphocytes in Werner syndrome patients

Summary The frequency of spontaneous 6-thioguanine (TG)-resistant peripheral blood lymphocytes in five unrelated Werner syndrome (WS) patients was determined using an autoradiographic labeling assay. The average frequency of TG-resistant lymphocytes was eightfold higher in WS patients than in sex- and age-matched normal control donors. This finding and previous identification of increased spontaneous chromosomal rearrangements and deletions in WS cells or cell lines suggest that WS is a human genomic instability or mutator syndrome.     

Clarification of chromosomal abnormalities associated with sexual ambiguity by studies with Y-chromosomal DNA sequences

Cases of gonadal dysgenesis, both Turner syndrome and mixed, were studied with Y centromeric and short-arm probes. The Y-centromeric alphoid repeat clone, Y97, allowed sensitive detection of Y-chromosomal material in marker chromosomes or mosaics by in situ analysis or Southern hybridization with purified DNA. The Y short-arm probe, p75/79, allowed detection of sequences normally associated with proximal Yp by Southern analysis. The presence of DNA fragments characteristic of Yp correlates well with partial male sexual differentiation in the cases of mixed gonadal dysgenesis. Thus, the combined use of molecular and cytogenetic techniques has proven to be a powerful approach to the analysis of chromosomal sex disorders.     

Intermediate range effects in DNA. I: Low pH/stress induced conformational changes in the vicinity of an extruded d(AT)n.d(AT) in cruciform.

A family of plasmids which contain d(AT)n cruciforms are sensitive to "single strand specific" (SS) endonucleases and a variety of chemical probes in a "random sequence" region centered 10-30 residues away from the cruciform junction. The SS nuclease sensitive structures are dependent on the presence of the extruded cruciform and exhibit a degree of sequence independence. Their appearance depends upon the combined effects of slightly lower than neutral pH and superhelical coiling above the minimum required to drive the extrusion of the d(AT)n cruciform arms. The nuclease sensitive structure is therefore underwound with respect to the B conformation and contains protonated bases.     

Nucleic acid studies on some heterofermentative lactobacilli: Description of Lactobacillus malefermentans sp.nov. and Lactobacillus parabuchneri sp.nov.

Chemotaxonomic studies were performed on some heterofermentative lactobacilli of uncertain taxonomic position. Two strains from beer and six strains from a variety of habitats were found to be distinct from each other and all other Lactobacillus species examined on the basis of DNA-DNA hybridizations and warrant new species for which the names L. malefermentans and L. parabuchneri , respectively, are proposed.     

The osmotic/calcium stress theory of brain damage: Are free radicals involved?

This overview presents data showing that glucose use increases and that excitatory amino acids (i.e., glutamate, aspartate), taurine and ascorbate increase in the extracellular fluid during seizures. During the cellular hyperactive state taurine appears to serve as an osmoregulator and ascorbate may serve as either an antioxidant or as a pro-oxidant. Finally, a unifying hypothesis is given for seizure-induced brain damage. This unifying hypothesis states that during seizures there is a release of excitatory amino acids which act on glutamatergic receptors, increasing neuronal activity and thereby increasing glucose use. This hyperactivity of cells causes an influx, of calcium (i.e. calcium stress) and water movements (i.e., osmotic stress) into the cells that culminate in brain damage mediated by reactive oxygen species.Special issue dedicated to Dr. Frederick E. Samson