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C Dickson

Publications and source records attributed to C Dickson.

At least 109 records · Page 6Linked to original sources

Multiple RNAs expressed from the int-2 gene in mouse embryonal carcinoma cell lines encode a protein with homology to fibroblast growth factors.

Mouse embryonal carcinoma cell lines that differ in their patterns of expression of the potential oncogene int-2 have been exploited in a structural analysis of the multiple RNA transcripts characteristic of this gene. Ribonuclease protection experiments indicate that four major classes of int-2 RNA initiate at heterogeneous cap sites within two distinct promoter regions, P1 and P2, spanning approximately 50 and 150 bp respectively. The more downstream promoter P2 is located in a region of the DNA that constitutes an intron in RNA transcripts that initiate at the upstream promoter P1. Otherwise, all four RNA structures share the same splice donor and acceptor sites that define the boundaries of the second and third exons. Further complexity arises through usage of two distinct polyadenylation signals, both variants of the normal consensus, that are separated by 1100 bp. Despite these structural variations, the results suggest that all four major classes of RNA encode the same protein product which shows significant homology to the family of heparin-binding proteins typified by basic fibroblast growth factor (FGF).

Animals↗

Amplification of the int-2 gene in primary human breast tumors.

Proviral activation of the int-2 gene is a frequent occurrence in mammary carcinomas induced by mouse mammary tumor virus (MMTV). Here, we have examined the human homolog of int-2 in 110 primary breast cancer DNAs. The locus was amplified 2- to 15-fold in 18 of the tumor DNA samples. Amplification of int-2 has a highly significant association (P less than 2 X 10(-6] with tumors from patients who subsequently developed a local recurrence of the disease or a distal metastasis.

Animals↗

Complete nucleotide sequence of a milk-transmitted mouse mammary tumor virus: two frameshift suppression events are required for translation of gag and pol.

We sequenced two recombinant DNA clones constituting a single provirus of the milk-transmitted mouse mammary tumor virus characteristic of BR6 mice. The complete provirus is 9,901 base pairs long, flanked by 6 base-pair duplications of cellular DNA at the site of integration. Five extensive blocks of open reading frame corresponding to the gag gene, the presumed protease, the pol and env genes, and the open reading frame orf within the long terminal repeat of the provirus were readily discernible. Translation of gag, protease, and pol involved three different translational reading frames to produce the three overlapping polyprotein precursors Pr77, Pr110, and Pr160 found in virus-infected cells. Synthesis of the reverse transcriptase and endonuclease therefore required two separate frameshifts to suppress the termination codons at the ends of the Pr77 and Pr110 domains. Direct evidence is presented for translational readthrough of both stop codons in an in vitro protein synthesis system.

Amino Acid Sequence↗

Insertion elements and transitions in cloned mouse mammary tumour virus DNA: further delineation of the poison sequences.

The provirus of mouse mammary tumour virus (MMTV) is reputed to contain sequences within the viral gag gene that prevent or inhibit its propagation as a recombinant DNA clone in Escherichia coli. Here we report the successful isolation of several lambda and plasmid clones comprising the 5' virus-host DNA junction fragments from integrated MMTV proviruses in BR6 mice. Although the lambda clones appeared intact, almost all of the plasmids were found to contain the bacterial insertion sequences IS1 or IS2 within a small region of the gag gene. One nondisrupted clone was recovered which had undergone multiple G to A transitions, some of which created stop codons in gag. These results have provided more precise information as to the location of the poison sequences and are discussed in relation to possible explanations for the phenomenon.

Amino Acid Sequence↗

Sequence, topography and protein coding potential of mouse int-2: a putative oncogene activated by mouse mammary tumour virus.

A major proportion of carcinomas induced by mouse mammary tumour virus (MMTV) show evidence for proviral activation of a cellular gene, int-2, on chromosome 7. The sequence of 7869 bp of DNA spanning the transcription unit of int-2 was determined and compared with that of a series of int-2-specific cDNA clones derived from mammary tumour RNA. The predicted positions of intron-exon boundaries, established by alignment of cDNA and chromosomal DNA sequences, indicate that the gene comprises at least three exons. An open reading frame capable of encoding a protein of 245 amino acids with an estimated mol. wt of 27 kd, is flanked by substantial non-coding segments at both 5' and 3' ends. Comparison of the chromosomal DNA sequence and the predicted amino acid sequence with available data-bases has revealed no homology to other known genes. These results are discussed in relation to the status of int-2 as a candidate proto-oncogene.

Amino Acid Sequence↗

Characterization, chromosome assignment, and segregation analysis of endogenous proviral units of mouse mammary tumor virus.

In the course of analyzing sites of proviral integration in tumors induced by mouse mammary tumor virus (MMTV), we have isolated recombinant DNA clones corresponding to the 5' and 3' ends of four endogenous MMTV proviruses present in BALB/c and BR6 mice. This has permitted the structural characterization of each locus by detailed restriction mapping and the preparation of DNA probes specific for the cellular sequences flanking each provirus. These probes have been used to trace the segregation patterns of the proviruses, designated Mtv-8, Mtv-9, Mtv-17, and Mtv-21, in a panel of inbred strains of laboratory mice and to map Mtv-17 and Mtv-21 to mouse chromosomes 4 and 8, respectively. The unambiguous resolution of these four proviruses on Southern blots has greatly facilitated the analysis of other endogenous MMTV proviruses in these inbred mice.

Animals↗

Characterization and chromosome assignment of the human homolog of int-2, a potential proto-oncogene.

int-2 is one of two cellular genes (int-1 and int-2) currently implicated in the genesis of mammary carcinomas by mouse mammary tumor virus and may constitute a novel cellular proto-oncogene. Using low-stringency hybridization with mouse int-2 probes, we established that homologous genes exist in a variety of mammalian species, including humans, but failed to detect related sequences in other classes and phyla. Recombinant bacteriophage clones and a single cosmid encompassing the human int-2 gene were isolated and characterized by restriction enzyme mapping. A survey of nine primary human breast tumors, three breast tumor cell lines, and three normal individuals revealed no evidence for gross amplification or rearrangement of the int-2 locus. Three distinct restriction fragment length polymorphisms were observed which could prove useful in future linkage studies. By a combination of in situ hybridization of metaphase chromosomes and somatic cell genetics, the human int-2 gene was mapped to chromosome 11, band q13.

Animals↗

Evidence for the involvement of descending noradrenergic pathways in the antinociceptive effect of baclofen.

The role of descending monoaminergic pathways in the antinociceptive effect of baclofen following intraperitoneal (i.p.) administration was investigated by determining the effect of intrathecal (i.t.) administration of neurotoxins [6-hydroxydopamine (6-OHDA) and 5,6-dihydroxytryptamine (5,6-DHT)] and receptor antagonists (phentolamine and methysergide) on baclofen in the tail-flick and hot-plate tests. Pretreatment with 6-OHDA (20 and 50 micrograms) inhibited baclofen antinociception in both tests 4-13 days after treatment. but pretreatment with 5,6-DHT (20 and 100 micrograms) produced a slight increase in the tail-flick test. The higher doses of both neurotoxins produced hyperalgesia in the tail-flick test. In other experiments, baclofen was injected intraperitoneally followed by i.t. amine antagonists when a plateau level of antinociception was attained. Phentolamine (30-100 micrograms) reversed baclofen antinociception in both the tail-flick and hot-plate tests. Methysergide (30-100 micrograms) only reversed the effect of baclofen in doses greater than or equal to 50 micrograms in the tail-flick test. Phentolamine (15 and 30 micrograms) and methysergide (50 micrograms) antagonized the antinociceptive effect of i.t. noradrenaline. Both phentolamine and methysergide produced hyperalgesia in the tail-flick test. However, hyperalgesia per se does not appear to be the only factor responsible for the reversal of baclofen antinociception because dose and agent dissociations between these effects were observed. These results suggest that a major mechanism of action of baclofen in producing antinociception is the activation of noradrenergic pathways descending to the spinal cord.

5,6-Dihydroxytryptamine↗

Changes in colonic motility during the development of chronic large bowel obstruction.

The effect of chronic progressive distal large bowel obstruction on colonic motility was studied in six mini-pigs. Motility was detected in vivo during the development of obstruction with chronically implanted Ag/AgCl electrodes using an impedance technique. When the obstruction was complete the segments of bowel were resected and spontaneous contractile activity and response to cholinergic stimulation were studied in an organ bath. Any hypermotility resulting from obstruction was shortlived and a gradual state of hypomotility supervened proximal to the obstruction. Decompression of the bowel did not result in the immediate return of motility and the resected bowel was unresponsive to carbachol. These results suggest that spontaneous resolution of large bowel obstruction is unlikely and that motility disturbances are unlikely to be a cause of anastomotic dehiscence.

Animals↗

D-Baclofen is an antagonist at baclofen receptors mediating antinociception in the spinal cord.

The antagonistic action of D-baclofen at baclofen receptors mediating antinociception in the spinal cord was examined. Drugs were administered intrathecally to rats and effects on nociceptive threshold evaluated in the tail flick test. L-Baclofen, D-baclofen and the racemate produced dose-related increases in tail flick latency, with L-baclofen being twice as potent as the racemate and approximately 100 times more potent than D-baclofen. When D-baclofen was injected 15 min prior to L-baclofen, it produced a dose-related inhibition of the effect of L-baclofen. Concomitant administration produced a more ambiguous effect. Antagonism appeared specific for baclofen receptors because analogues with full and partial agonist activity as well as an agonist dose of D-baclofen, but not morphine or noradrenaline, were inhibited by pretreatment with D-baclofen. gamma-Aminobutyric acid (GABA) did not increase tail flick latency either alone or following pretreatment with an uptake inhibitor or a GABA-transaminase inhibitor. Antinociception produced by intrathecal administration of Baclofen appears to result from activation of a receptor which is stereoselective for the L-isomer and can be blocked by D-baclofen in doses which have initial agonist activity. This receptor may not be a GABA subtype because GABA does not mimic the effect of baclofen and the rank order of potency of analogues differs from established GABAB systems.

Alkynes↗

Changes in intestinal blood flow during the development of chronic large bowel obstruction.

The effect of chronic progressive distal large bowel obstruction on intestinal blood flow was studied in 12 mini-pigs. Blood flow was measured by the techniques of xenon133 clearance and intracardiac injections of radiolabelled microspheres. Intestinal blood flow was increased in the left colon and ileum but decreased in the caecum. Blood was shunted from the mucosa to the muscle layers. The results suggest that primary anastomosis of the obstructed left colon is not contraindicated by haemodynamic considerations.

Animals↗

Mouse mammary tumor virus can mediate cell fusion at reduced pH.

Mouse mammary tumor virus, a type-B retrovirus, was shown to mediate fusion of cultured cells following low-pH treatment. Fusion could be demonstrated both with virus-infected cells or with uninfected cells carrying freshly absorbed virus. Although the fusion response was variable between different cell lines, one line of MMTV-infected mink lung cells, designated MGR4, was particularly susceptible to fusion at reduced pH. Since expression of MMTV in these cells is strongly regulated by glucocorticoids, it was possible to demonstrate that cell fusion was dependent on MMTV-encoded functions. With MGR4 cells, a pH threshold for membrane fusion was observed, centered on pH 5.5, at which 50% of the cells were fused. At lower pHs virtually all of the cells in the monolayer fused. These results are similar to those described for other virus groups and are consistent with the idea that most enveloped animal viruses infect cells by a common mechanism involving membrane fusion triggered by low pH.

Animals↗