Constitutive biosynthesis of bone Gla protein in a human osteosarcoma cell line.

A human osteosarcoma cell line derived from cells obtained from a patient with Paget's disease is shown to synthesize and secrete bone Gla protein (BGP); (osteocalcin), a noncollagenous bone matrix protein. Using a human BGP-specific RIA, we show that the human osteosarcoma cells synthesize significant amounts of BGP without any prior induction of BGP synthesis by 1,25-dihydroxyvitamin D. After specific immunoprecipitation of poly-A+ RNA in vitro translation products with antibodies to BGP, we found that BGP is synthesized as a precursor with an apparent mol wt of 13.5K, as demonstrated on 15% sodium dodecyl sulfate-polyacrylamide gels. Finally, pulse labeling of the osteosarcoma cells with [3H]proline reveals that the cells synthesize mature BGP of 12,000 mol wt as well as a higher mol wt precursor (13,500) of the protein.

Telescience at the University of California, Berkeley.

The University of California at Berkeley (UCB) is a member of a university consortium involved in telescience testbed activities under the sponsorship of NASA. Our Telescience Testbed Project consists of three experiments using flight hardware being developed for the Extreme Ultraviolet Explorer project at UCB's Space Sciences Laboratory. The first one is a teleoperation experiment investigating remote instrument control using a computer network such as the Internet. The second experiment is an effort to develop a system for operation of a network of remote workstations allowing coordinated software development, evaluation, and use by widely dispersed groups. The final experiment concerns simulation as a method to facilitate the concurrent development of instrument hardware and support software. We describe our progress in these areas.

Analysis of DNAs from two species of the virilis group of Drosophila and implications for satellite DNA evolution.

The DNAs from two virilis group species of Drosophila, D. lummei and D. kanekoi, have been analyzed. D. lummei DNA has a major satellite which, on the basis of CsCl equilibrium centrifugation, thermal denaturation, renaturation and in situ hybridization is identical to D. virilis satellite I. D. kanekoi DNA has a major satellite at the same buoyant density in neutral CsCl gradients as satellite III of D. virilis. However, on the basis of alkaline CsCl gradients, the satellite contains a major and a minor component, neither one of which is identical to D. virilis satellite III. By in situ hybridization experiments, sequences complementary to the major component of the D. kanekoi satellite are detected in only some species and in a way not consistent with the phylogeny of the group. However, by filter hybridization experiments using nick-translated D. kanekoi satellite as well as D. lummei satellite I and D. virilis satellite III DNAs as probes, homologous sequences are detected in the DNAs of all virilis group species. Surprisingly, sequences homologous to these satellite DNAs are detected in DNAs from non-virilis group Drosophila species as well as from yeast, sea urchin, Xenopus and mouse.

Insulin receptor overexpression in a human pre-B acute lymphocytic leukemia cell line with a t(1;19) chromosome translocation near the INSR locus.

Human pre-B acute lymphocytic leukemia (ALL) cell line 697 contains a disease-specific chromosomal translocation, t(1;19), near the insulin receptor locus (INSR). Insulin binding, insulin receptor kinase and cell surface immunofluorescence experiments all show that 697 cells express more surface insulin receptors than 207 cells, a line of pre-B ALL cells that lack the t(1;19). Northern blot analysis confirms that 697 cells have increased levels of mRNA for the insulin receptor. Gene dosage and in situ hybridization analysis indicate that only two copies of the INSR locus are present in 697 cells, one on the translocation chromosome and one on the normal chromosome 19. The results described here demonstrate that transcriptional up-regulation of the INSR locus is correlated with the t(1;19) translocation found in human pre-B ALL cell line 697.

Cystic fibrosis DNA markers in Alabama blacks.

DNA samples from unrelated Alabama blacks with no family history of cystic fibrosis (CF) were analyzed with DNA markers linked to cf. Allelic frequencies of genetic markers detected by probes pmetH, pmetD, XV2c, KM19, and pJ3.11 were compared to those of other populations. Allelic frequencies for pJ3.11, XV2c, and KM19 in Alabama blacks and previously studied Caucasian populations were similar. In contrast, the met locus allelic frequencies in Alabama blacks were markedly different from those in Caucasian populations.

Complete detection of mutations in cystic fibrosis patients of Native American origin.

An increased incidence of cystic fibrosis (CF) has been reported in some populations of Native Americans of the Southwest such as the Pueblo, which is a genetic isolate. As the most common mutation found in Caucasians (delta F508) was absent and only one chromosome carried the G542X mutation, we decided to analyze the entire coding sequence of the CFTR gene in eight Pueblo CF patients. We have identified four different mutations: G542X, R1162X, 3849+10kbC-->T, and D648V that account for these 16 haplotypes. The R1162X was found on 11 chromosomes. Using intragenic microsatellites, we have compared the haplotypes of those chromosomes to those of Italian origin where the R1162X mutation was initially reported. These haplotypes turned out to be identical, suggesting a common origin and an admixture with Italian or Spanish settlers, followed by typical founder effect. In contrast the 3849+10kbC-->T mutation, which was found on three chromosomes, is associated with different haplotypes than those on chromosomes carrying the same mutation in Caucasians. A novel mutation, D648V, observed on one chromosome has not been found outside the Pueblo population.