Predominance of null mutations in ataxia-telangiectasia

Ataxia-telangiectasia (A-T) is an autosomal recessive disorder involving cerebellar degeneration, immunodeficiency, chromosomal instability, radiosensitivity and cancer predisposition. The responsible gene, ATM, was recently identified by positional cloning and found to encode a putative 350 kDa protein with a PI 3-kinase-like domain, presumably involved in mediating cell cycle arrest in response to radiation-induced DNA damage. The nature and location of A-T mutations should provide insight into the function of the ATM protein and the molecular basis of this pleiotropic disease. Of 44 A-T mutations identified by us to date, 39 (89%) are expected to inactivate the ATM protein by truncating it, by abolishing correct initiation or termination of translation, or by deleting large segments. Additional mutations are four smaller in-frame deletions and insertions, and one substitution of a highly conserved amino acid at the PI 3-kinase domain. The emerging profile of mutations causing A-T is thus dominated by those expected to completely inactivate the ATM protein. ATM mutations with milder effects may result in phenotypes related, but not identical, to A-T

Reconstructing the deep-branching relationships of the papilionoid legumes

Resolving the phylogenetic relationships of the deep nodes of papilionoid legumes (Papilionoideae) is essential to understanding the evolutionary history and diversification of this economically and ecologically important legume subfamily. The early-branching papilionoids include mostly Neotropical trees traditionally circumscribed in the tribes Sophoreae and Swartzieae. They are more highly diverse in floral morphology than other groups of Papilionoideae. For many years, phylogenetic analyses of the Papilionoideae could not clearly resolve the relation- ships of the early-branching lineages due to limited sampling. In the eight years since the publication of Legumes of the World, we have seen an extraordinary wealth of new molecular data for the study of Papilionoideae phylogeny, enabling increasingly greater resolution and many surprises. This study draws on recent molecular phylogenetic studies and a new comprehensive Bayesian phylogenetic analysis of 668 plastid matK sequences. The present matK phylogeny resolves the deep-branching relationships of the papilionoids with increased support for many clades, and suggests that taxonomic realignments of some genera and of numerous tribes are necessary. The potentially earliest-branching papilionoids fall within an ADA clade, which includes the recircumscribed monophyletic tribes Angylocalyceae, Dipterygeae, and Amburanae. The genera Aldina and Amphimas represent two of the nine main but as yet unresolved lineages comprising the large 50-kb inversion clade. The quinolizidine-alkaloid-accumulating Genistoid s.l. clade is expanded to include Dermatophyllum and a strongly supported and newly circumscribed tribe Ormosieae. Sophoreae and Swartzieae are dramatically reorganized so as to comprise mono-phyletic groups within the Core Genistoid clade and outside the 50-kb inversion clade, respectively. Acosmium is excluded from the Genistoids s.l. and strongly resolved within the newly circumscribed tribe Dalbergieae. By providing a better resolved phylogeny of the earliest-branching papilionoids, this study, in combination with other recent evidence, will lead to a more stable phylogenetic classification of the Papilionoideae.Web of Scienc

Effect of novobiocin on DNA synthesis and structure in human lymphoblastoid cells

Novobiocin has an inhibitory effect on DNA synthesis in human lymphoblastoid cells similar to that observed with other mammalian cells. A concentration and time-dependent increase in sedimentation of DNA nucleoids, derived from human lymphoblastoid cells, was observed in the presence of novobiocin. No decrease in the degree of negative supercoiling of DNA, determined by sedimentation on sucrose gradients in the presence of ethidium bromide, was obtained when cells were incubated with novobiocin. The latter findings are contrary to results obtained with other mammalian cells. Alteration in sedimentation rate of nucleoids after novobiocin treatment, at concentrations up to 0.25mg/ml, does not correlate with inhibition of DNA synthesis

Defective DNA topoisomerase II in ataxia-telangiectasia cells.

A number of characteristics in the human genetic disorder ataxia-telangiectasia are compatible with an alteration to chromatin structure or the recognition of that structure by an enzyme or DNA binding protein. We describe here reduce activity of DNA topoisomerase type II in a number of Epstein Barr Virus-transformed ataxia-telangiectasia lymphoblastoid cell lines. Enzyme activity was reduced 10-fold or greater in 4 out of 5 cell lines compared to controls. In the remaining cell line approximately a 2-3 fold reduction was evident in partially purified extracts. DNA topoisomerase type I activity was found to be the same as controls in all the cell lines. Northern blot analysis revealed that the same level of DNA topoisomerase II mRNA was expressed in ataxia-telangiectasia and control cell lines. The size and amount of the enzyme did not differ appreciably from that observed in control cells. The reduced activity of DNA topoisomerase II in ataxia-telangiectasis cells might be explained by amino acid substitutions, small deletions in DNA or by a defect in post-translational modification in these cells

p-Aminophenol induced DNA damage and cytotoxicity enhanced by autoxidation

p-Aminophenol inhibits DNA synthesis and alters the structure of DNA. A decrease in sedimentation of nucleoids from cells treated with p-aminophenol was observed and this decrease in sedimentation was considerably less when cells were incubated with p-aminophenol in an atmosphere of nitrogen or at lower pH values. This compound was also shown to be cytotoxic to cells in culture. These results demonstrate that conditions retarding the autoxidation of p-aminophenol lead to reduced effects on DNA structure and a lesser cytotoxic effect

Lymphocytes in ataxia telangiectasia: Implications of the DNA repair defect

Fibroblasts from patients with ataxia telangiectasia have been demonstrated to be more sensitive to ionizing radiation than controls and are defective in repair of such damage to DNA. We have investigated the DNA repair capacity of phytohemagglutinin-stimulated lymphocytes, from persons with ataxia telangiectasia, after exposure to ionizing radiation. Comparison with DNA repair in control lymphocytes indicates a much reduced capacity in ataxia telangiectasia. Measurement of DNA repair in stimulated lymphocytes thus provides a suitable and rapid assay for AT homozygotes. The lymphocyte DNA repair defect may be related to the immunological abnormalities seen in this disease

Internal tides in the Northern Gulf of California

The characteristics of the internal tide in the Northern Gulf of California are described using data from two moored arrays of temperature and current sensors, one for summer and one for winter, located between Angel de la Guarda Island and the mainland. From the summer six-sensor mooring it was found that: (1) the current fluctuations are dominated by the semidiurnal frequency band, while the quarterdiurnal frequency dominated the temperature fluctuations. (2) The baroclinic semidiurnal horizontal current fluctuations are aligned with the gulf axis, and have amplitudes of 10-15 cm s-1; the vertical displacements reached 4 m in this frequency band. (3) The vertical modal structure for the temperature and velocity oscillations was dominated by the first and third modes. (4) The energy of the semidiurnal internal tide is 45% of that of the barotropic tide. (5) Vertical wave number spectra showed slightly asymmetric peaks in the high wave number components, indicating that their downflowing energy is larger than that flowing upward. From the winter two-sensor mooring, it was found that the vertical oscillations were mainly semidiurnal, with root mean square amplitudes of 7 m

Inhibition of DNA, RNA and protein synthesis and chromatin alteration by N-hydroxyphenacetin

1. The effects of N-hydroxyphenacetin on DNA function and structure were investigated to elucidate the involvement of phenacetin in analgesic nephropathy and transitional cell carcinoma. 2. N-Hydroxyphenacetin or a metabolite inhibited synthesis of DNA, RNA and protein; DNA inhibition was greater at higher pH. 3. No single-strand breaks were detectable in DNA after N-hydroxyphenacetin treatment and no appreciable effect on cell viability was observed at concentrations up to 5mM. 4. N-Hydroxyphenacetin-induced alteration to chromatin structure was detected using nucleoid sedimentation analysis. Direct binding to plasmid DNA was not observed. 5. These observations are consistent with a role for phenacetin metabolites in renal disease