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D R Smyth

Publications and source records attributed to D R Smyth.

35 records · Page 2Linked to original sources
PubMed1989

An element with long terminal repeats and its variant arrangements in the genome of Lilium henryi.

A 9.35 kbp element with long terminal direct repeats (LTRs) of 2.4 kbp has been characterized from the large genome of Lilium henryi. The organization of the element, named del, was examined in 20 fragments from a genomic DNA library constructed using phage EMBL3. Five fragments apparently contained full 9.35 kbp elements while in 11 del was recovered in part. Two clones carried tandem arrangements of del, with single LTRs and internal sequences alternating, and one insert contained a solo LTR. Evidence suggests that these arrangements, which can arise by unequal crossing over, have a genomic rather than a cloning origin. Finally, one cloned fragment had a complex del arrangement yet to be fully defined. Limited sequencing of five LTRs (from a full del, a tandem arrangement and a solo LTR) indicates that the consensus termini are 5'TGT...ACA3', with del sequences flanked by a 5 bp tandem repeat. Thus del shares properties with retrotransposons such as the copia family of Drosophila and Ty of yeast. However with more than 13,000 copies per genome it seems del has been amplified more than is usual for such elements.

Base Sequence↗
PubMed1989

Lack of effect of maternal age on UV-induced DNA repair in mouse oocytes.

Oocytes at the dictyate stage from young (8-14 weeks) and old (12-15 months) BALB/c mice were manually isolated and UV-irradiated. They were cultured for 1 h in medium containing tritiated thymidine and chased for a further hour in cold thymidine medium before being incubated for 18-20 h in medium with no added thymidine. Oocytes which had developed to metaphase II were analysed following autoradiography. Pooled results from 14 replicate experiments revealed no significant age-related difference between the mean corrected grain count per cell [159.2 +/- 8.5 (86 cells) for young mice and 164.6 +/- 9.8 (70 cells) for the old animals]. Thus in the female mouse the oocyte's capacity to repair UV-induced damage is apparently maintained at a high level throughout reproductive life.

Animals↗
PubMed1989

Plant retrotransposon from Lilium henryi is related to Ty3 of yeast and the gypsy group of Drosophila.

The lily retrotransposon del 1-46 is 9345 base pairs (bp) long. It has long terminal repeats (LTRs) of 2406 bp (left) and 2415 bp (right), which differ in sequence by 1.4%. Sequences similar to those involved in priming DNA synthesis in retroviruses occur in the internal region. Near the left LTR is a sequence complementary to 18 residues at the 3' end of methionine initiator tRNA of three plant species, and a run of 12 purines occurs close to the right LTR. One internal reading frame of del 1-46 has relatively few stop codons. The 1462-codon product from this frame has motifs, in N to C terminus order, corresponding to those identified with RNA binding, protease, reverse transcriptase, RNase H, and integrase functions in retroviruses and certain other retrotransposons. Amino acid sequence comparisons of three conserved pol regions show del to be closely related to the Ty3 retrotransposon of yeast (37-40% identity). del is also related to the gypsy group of Drosophila (17.6, 297, gypsy/mdg4, and 412), showing closer identity with their reverse transcriptase (32-38%) and RNase H (36-45%) domains than with their integrase domain (21-26%). It is proposed that a gypsy group ancestor exchanged the integrase region with a more distantly related element since its divergence from a del/Ty3 common ancestor. The occurrence of related retrotransposons in three different kingdoms (plants, animals, and fungi) strongly implies their horizontal transmission in recent evolutionary time.

Amino Acid Sequence↗
PubMed1989

Genes directing flower development in Arabidopsis.

We describe the effects of four recessive homeotic mutations that specifically disrupt the development of flowers in Arabidopsis thaliana. Each of the recessive mutations affects the outcome of organ development, but not the location of organ primordia. Homeotic transformations observed are as follows. In agamous-1, stamens to petals; in apetala2-1, sepals to leaves and petals to staminoid petals; in apetala3-1, petals to sepals and stamens to carpels; in pistillata-1, petals to sepals. In addition, two of these mutations (ap2-1 and pi-1) result in loss of organs, and ag-1 causes the cells that would ordinarily form the gynoecium to differentiate as a flower. Two of the mutations are temperature-sensitive. Temperature shift experiments indicate that the wild-type AP2 gene product acts at the time of primordium initiation; the AP3 product is active later. It seems that the wild-type alleles of these four genes allow cells to determine their place in the developing flower and thus to differentiate appropriately. We propose that these genes may be involved in setting up or responding to concentric, overlapping fields within the flower primordium.

Alleles↗
PubMed1988

UV-induced DNA repair is not detectable in pre-dictyate oocytes of the mouse.

Oocytes from fetal and neonatal mice were UV-irradiated, cultured in medium containing tritiated thymidine and analysed following autoradiography. As previously reported irradiated dictyate cells clearly displayed an increased grain count. However we detected no UV-induced response in oocytes at the leptotene, zygotene or pachytene stages of meiosis. This contrasts with the situation in spermatogenesis where high levels of repair can be induced in equivalent early prophase stages.

Animals↗
PubMed1987

Silver bands in chronic granulocytic leukemia. II. The Philadelphia chromosome.

To test if rRNA gene activity in the Philadelphia (Ph) chromosome nucleolar organizing regions (NOR) differs from other NOR we scored silver banding patterns in leukemic cells of 32 patients (one patient was sampled twice) in the chronic phase of chronic granulocytic leukemia. The average number of bands per NOR in banded cells of these patients was 0.40 +/- 0.06 for the Ph NOR, not significantly different from the 0.41 +/- 0.02 per D chromosome NOR or the 0.41 +/- 0.04 per G. Across the 33 cases, the number of different Ph NOR banded in at least one cell (23 of 33, 70%) was also closely similar to the cumulative maximum number of chromosome D NOR banded per cell (137 of 198 banded, 69%) and G NOR banded (67 of 99, 67%). We conclude that the activity of rRNA genes in Ph NOR is closely similar to that of the other nine NOR in each CGL cell. In situ hybridization of 3H-labeled complementary rRNA to NOR of two patients revealed that the mean number of Ph NOR grains fell within the range of the other NOR. Also, grain distributions across the NOR of leukemic cells closely matched those of stimulated lymphocytes. Thus, a limited sampling has shown no evidence for consistent differences or changes in the rRNA gene number of Ph NOR.

Adult↗
PubMed1984

Silver bands in chronic granulocytic leukemia: I. Increased banding associated with blastic transformation.

We have tested the proposal that the level of silver banding in leukemic cells of Ph1 + chronic granulocytic leukemia (CGL) patients increases as the disease progresses. Blood and/or bone marrow cells from 14 patients were cultured for 24 hr before banding. In all but one case, there were two populations of mitoses, those with silver bands on their nucleolar organizing regions (NORs) and those without. The percentage of cells that banded was higher, on average, in cultures from 7 patients in blastic transformation (80%) than in 8 chronic cases (36%) or in one accelerated phase (49%). Also, the mean number of NORs stained in banded cells was higher in blastic phase (6.9) compared with chronic phase cells (4.4). Hyperdiploid cell lines were present in four cases of myeloblastic transformation. All such cells were silver banded, and the mean fraction of NORs banded in them was relatively high. An increase in silver banding with time was shown in two of the patients. It seems that silver banding does increase in CGL cells as the disease progresses. This may arise either through an increase in the rate of ribosomal RNA synthesis in leukemic cells present in the blastic phase or possibly by a decrease in the rate of degradation (or processing) of newly synthesized rRNA.

Adult↗
PubMed1982

Patterns of exchange induced by mitomycin C in C-bands of human chromosomes. I. Relationship to C-band size in chromosomes 1, 9, and 16.

Frequencies of exchange were determined in C-bands of chromosomes 1, 9 and 16 in six normal males, and related to relative C-band area. Comparing these different chromosomes, more exchanges occurred on average in 9 than in 1 although their mean C-band sizes were similar. Chromosome 16 exchanges were fewer, both overall and relative to C-band area. Comparing the same chromosome between individuals, there was a positive correlation between relative frequency and band size in both 1-1 and 9-9 exchanges. No clear trend was observed for other exchange events. If homology is required for interchange, it cannot be dependent solely on overall C-band size. Perhaps certain DNA sequences, sensitive to mitomycin C damage, are located in part of each C-band, with less per unit area in chromosome 1 than in 9 and still less in chromosome 16. X- and U-type exchanges between chromosome 9s occurred in near equal frequencies in all individuals. If synapsis of specific, affected sequences is a pre-requisite for interchange, this observation suggests that the affected sequence in chromosome 9 is arranged in both orientations relative to the centromere.

Chromosome Banding↗
PubMed1982

Patterns of exchange induced by mitomycin C in C-bands of human chromosomes. II. High frequency of Y-Y exchange in XYY cells.

In XY cells, exchanges in the Y C-band were rare (less than 0.8%). However, in XYY cells we observed 58 Y chromosomes involved in exchange out of 208 (28%). Most were Y-Y interchanges. Therefore mitomycin C does affect Y C-bands, but subsequent interchange requires the presence of a homologue. This may be the consequence of interphase association of the homologues, or by homologous interaction of an affected DNA sequence in the two Ys. If the latter, the proposed sequence cannot be localized to proximal or distal parts of the Y C-band, as exchange points were observed in both. Also, it may be oriented in one direction in the Y, as the Y-Y exchanges which could be unambiguously classified were X-type events.

Chromosome Banding↗
PubMed1981

DNA extraction during Giemsa differentiation of chromatids singly and doubly substituted with BrdU.

Human lymphocytes were cultured in 3H-labelled BrdU. Cells were pretreated to induce differentiation, autoradiographed and Giemsa-stained. DNA extraction was deduced if grain counts were lower in differentiated mitoses compared with untreated controls.--The differentiation method involved sequential pretreatments with short wave UV and 2 x SSc at 60 degrees C. This removed 34% of label from first division cells (with TB.TB chromosomes) but relatively more (53%) from second division (TB.BB chromosomes). In second division cells, about two thirds of label was lost from pale (BB) chromatids but only one third from dark (TB) chromatids. The UV and SSC pretreatments acted in collaboration, since neither alone reduced grain counts significantly.--On testing other methods, similar preferential DNA extraction was obtained with Perry and Wolff's FPG method, and with the hot salt pretreatment of Korenberg and Freedlender. However, good Giemsa differentiation could also be obtained using Hoechst 33258 and light pretreatments without any DNA loss. Reverse differentiation patterns (TB pale, BB dark) induced by warm acids resulted in extraction of nearly two thirds of 3H-BrdU label, but relative loss was the same from pale and dark chromatin. Direct reverse staining using alkaline Giemsa did not result in any loss of label.--Thus preferential DNA loss from pale stained chromatin underlies differentiation methods using light plus hot salt pretreatments, but it is not obligatory for good differentiation using other techniques.

Autoradiography↗
PubMed1976

Mapping of sister-chromatid exchanges in human chromosomes using G-banding and autoradiography.

Lumphocytes were pulse-labelled with [3H] thymidine. Following G-banding, the cells were autoradiographed and 46 in their third post-labelling division selected. The locations of 611 sister-chromatid exchanges (SCE's) which had occurred in the previous two cell cycles were recorded as label discontinuities along identified chromosomes. Between particular chromosomes, SCE frequency was proportional to chromosome length. SCE frequency distributions within particular chromosomes fitted Poisson expectations. There was no over-representation of exchanges in centromeric regions, or in the C-banded regions of chromosomes 1, 9 and 16. A trend of increased frequency of SCE in darkly G-banded regions and in relatively darkly banded chromosomes was evident. The apparent excess of SCE in dark G-bands could be considered to be a consequence of the more condensed state of the DNA in these regions in the interphase nucleus relative to the DNA in pale G-band regions. Such compaction could result in an enhanced probability of SCE and a reduced probability of gross inter- or intra-change involving these regions. In contrast, the more extended interphase state of the DNA in pale G-banded regions would allow non-homologous exchange and account for the preferred location of X-ray-induced exchange events to pale G-bands.

Autoradiography↗