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P R Harrison

Publications and source records attributed to P R Harrison.

At least 55 records · Page 3Linked to original sources
PubMed1986

The structure of the mouse glutathione peroxidase gene: the selenocysteine in the active site is encoded by the 'termination' codon, TGA.

Glutathione peroxidase (GSHPx) is an important selenium-containing enzyme which protects cells from peroxide damage and also has a role in leukotriene formation. We report the identification of a genomic recombinant as encoding the entire mouse GSHPx gene. Surprisingly, the selenocysteine in the active site of the enzyme is encoded by TGA: this has been confirmed by primer extension/dideoxy sequencing experiments using reticulocyte mRNA. The same site of transcription initiation is used in three tissues in which the GSHPx mRNA is expressed at high levels (erythroblast, liver and kidney). Like some other regulated 'house-keeping' genes, the GSHPx gene has Sp1 binding site consensus sequences but no 'ATA' and 'CAAT' consensus sequences upstream of the transcription initiation site. Moreover, there is a cluster of two Sp1 binding site consensus sequences and two SV40 core enhancer sequences in the 3' region of the gene, close to the previously mapped position of a DNase I-hypersensitive site found only in tissues expressing the GSHPx mRNA at high levels.

Amino Acid Sequence↗
PubMed1985

Analysis of chromatin changes associated with the expression of globin and non-globin genes in cell hybrids between erythroid and other cells.

Red blood cell differentiation involves the coordinate expression of a set of polypeptides some of which are erythroid-specific (the abundant globins as well as minor species such as glycophorin, carbonic anhydrase I and the RBC lipoxygenase) whereas others are found also in a subset of other cells, e.g. beta spectrin and a 19 kd polypeptide (ep 19) found in adult liver and kidney as well as erythroid cells. To investigate the genetic mechanisms involved in the regulation of these classes of genes, the expression of lipoxygenase, ep 19 and beta globin mRNAs was investigated in cell hybrids between mouse erythroid (Friend) cells and mouse T-lymphoma or neuroblastoma cells. All three mRNAs are expressed or repressed together in cell hybrids between the Friend cell and lymphoma or neuroblastoma cells respectively. Moreover, studies of the chromatin structure surrounding the genes reveal that erythroid cell-specific DNaseI hypersensitive sites within the ep 19 and beta major globin genes are lost in the Friend cell X neuroblastoma hybrids whereas they are retained in the Friend cell X lymphoma cell hybrids. This implies that the trans-acting mechanism responsible for regulating the RBC phenotype in these cell hybrids acts at the level of the early chromatin changes thought to reflect a pre-activation stage in gene expression.

Animals↗
PubMed1984

Molecular analysis of erythropoiesis. A current appraisal.

This article considers recent evidence concerning the molecular mechanisms involved in the coordinate regulation of gene expression during red blood cell (RBC) differentiation. Contrary to popular belief, recent evidence shows that only a few of the characteristic RBC proteins are restricted to the erythroid lineage: apart from the globins, an RBC lipoxygenase and (possibly) glycophorin are the only examples for which there is reasonably good evidence. In contrast, the proteins forming the RBC cytoskeleton (spectrin, ankyrin, band 4.1, actin and possibly the major anion exchange transmembrane protein by which the cytoskeleton is attached to the plasma membrane) have closely-related variants in other cell types. Yet two beta-spectrin variants are found exclusively in certain terminally differentiated cells, often only in certain specific regions of the cell membrane. Certain RBC isozymes (e.g. for pyruvate kinase and carbonic anhydrase) and an RBC 19 kD protein (ep19) are also expressed only in a subset of other cell types. This illustrates the importance of gene families which are differentially regulated in certain subsets of cell types during differentiation and development. The expression of the globin genes seems to be regulated mainly at the transcriptional level, although transport of these transcripts to the cytoplasm may be controlled by interactions with other RNAs: stabilisation of globin mRNAs by ribonucleoprotein complexes in the cytoplasm may also be important. In fact, the expression of the globin genes involves two distinct phases: first, structural changes occur in the chromatin surrounding the genes (as determined by sensitivity to digestion by nucleases) and these can be maintained independently of any subsequent transcription. In many cases, these nuclease-sensitive sites in the chromatin correspond to low-level transcription initiation sites and to DNA sequences with regulatory functions when the isolated genes are assayed for transcription in vivo after transfection into cells. How the unlinked alpha- and beta-globin genes are coordinately regulated is not yet understood. Indeed, the alpha- and beta-gene promoters have quite different properties as judged by their responses to DNA replication and to factors known to affect viral gene function (e.g. the cis-acting SV40 enhancer elements and the trans-acting adenovirus regulatory protein, Ela). Other evidence shows that a nuclear protein present only in erythroid cells is able to bind to the beta-globin gene precisely in the region that is hypersensitive to nuclease digestion in chromatin from erythroid cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1984

Glomerulonephritis, non-Hodgkin's lymphoma and leucocytoclastic vasculitis.

2 cases of active focal proliferative glomerulonephritis, lymphocytic malignant lymphoma and leucocytoclastic vasculitis are described. 1 presented with the nephrotic syndrome and progressive renal impairment, the other with acute renal failure. Cryoglobulins were detected in the serum of 1 patient, but not in that of the other. Initial renal and skin biopsies in the 2 patients were strikingly similar. As well as demonstrating glomerular lesions, the renal biopsies contained interstitial lymphomatous infiltrate which, in 1 case, was initially incorrectly interpreted and resulted in delay in specific treatment. Immunosuppressive therapy markedly improved renal function and induced remission of the vasculitis and lymphoma. Repeat renal biopsy in 1 case confirmed regression of the glomerulonephritis. A common immunological aetiology may explain the co-existence of these three conditions in the 2 patients.

Biopsy↗
PubMed1983

Isolation of non-globin genes expressed preferentially in mouse erythroid cells.

Genomic DNA recombinants were isolated from a library of Balb-C mouse genomic DNA fragments cloned in lambda Ch4A by screening with cDNA derived from 13d foetal liver cell or adult reticulocyte poly A+ RNA. Subsequent screening enabled us to identify non-globin genomic sequences whose expression appeared exclusive to or elevated in erythroid cells. Further analysis of the structure and expression of these sequences was performed using Southern blot and DNA or RNA dot hybridisation analysis. In one recombinant part of the cloned genomic sequence corresponded to an erythroblast specific mRNA identified previously by Affara et al, (5).

Animals↗
PubMed1983

Patterns of expression of erythroblast non-globin mRNAs.

Three classes of erythroblast non-globin mRNAs have been identified using cDNA recombinants isolated from a mouse foetal liver cDNA library. One erythroid cell-specific 12S mRNA coding for a 16-19,000 dalton protein has been identified using two independent cDNA recombinants (pFC5 and pFA6). The gene encoding this mRNA is unique in the genome and its organisation seems to be the same in both erythroid and non erythroid cell genomic DNA as judged by digestion with restriction enzymes and Southern blotting. Another erythroblast mRNA of size 16S represented by recombinant pFD12 is expressed in brain cells as well as a variety of haemopoietic cell types, but not in adult liver or fibroblasts. Two other erythroblast mRNAs of size 8S and 12S represented by recombinants pD6 and pA4 are expressed in many differentiated cell types with the exception of non-dividing cells such as reticulocytes, peripheral white blood cells, adult liver and brain cells. These mRNAs are, therefore, presumed to be involved ubiquitously in cell proliferation or general cell metabolism.

Animals↗
PubMed1983

Long-term effects of endralazine (BQ-22-708) in patients with renal impairment and hypertension.

Endralazine, a new peripheral vasodilator, was studied in 21 patients with hypertension and chronic renal failure. Nineteen patients had unacceptable control of hypertension with their previous therapy, and 2 were suffering adverse effects for other third line drugs. All patients continued to receive a beta-adrenergic blocking agent. Five patients failed to complete the study, two because of poor compliance, and 3 as a result of failure to control raised blood pressure. In the remaining 16 patients, satisfactory blood pressure reduction was seen at 6 months and was maintained in 12 patients followed for 18 months. Endralazine was well tolerated. No patient developed the lupus syndrome nor evidence of drug-induced immunological abnormality.

Adult↗
PubMed1981

Cloning of a new mouse foetal beta-globin mRNA sequence.

A novel globin cDNA recombinant (pFG5) has been isolated from a 14-15 day Porton mouse foetal liver cDNA library. It codes for a beta-like globin mRNA expressed in foetal liver-derived erythroblasts and erythrocytes but not in adult reticulocytes nor in yolk sac derived nucleated erythrocytes. It is also found in Friend cells induced to differentiate by DMSO. The nucleotide sequence of pFG5 confirms that it does not code for the beta major or beta minor globin chains nor the embryonic epsilon Y2 globin chain; but it is identical to the published partial sequence of the epsilon Y3 globin gene over the region of overlap (78 nucleotides).

Animals↗
PubMed1981

Transfer and expression of herpes simplex virus thymidine kinase and human globin genes in mammalian cells studied by spot hybridization assays.

A spot hybridization assay has been developed for rapid estimations of specific DNA sequences in whole mammalian cells or DNA. This method is applicable to gene-transfer studies in determining the copy number(s) of foreign DNA(s) introduced into mammalian cells. A similar assay has been developed for determining amounts of specific RNA sequences in mammalian cells. It is also shown that this method is applicable to gene-transfer studies and in particular to determining the expression of foreign genes introduced into cells.

Animals↗
PubMed1981

Cell-cycle dependence of induced hemoglobin synthesis in Friend erythroleukemia cells temperature-sensitive for growth.

Friend erythroleukemia cells temperature-sensitive (ts) for growth have been used to study the cell-cycle dependence of erythroid differentiation. ts cells accumulated hemoglobin in response to inducer when replicating optimally at permissive temperatures. However, when arrested in the G1 phase of the cell cycle at a nonpermissive temperature, ts cells did not accumulate hemoglobin or globin mRNA nor did they differentiate terminally. Under these conditions, the viability of ts cells treated with inducer at the nonpermissive temperature remained high (70% of that of cells induced at the permissive temperature) as judged by cloning experiments. Total RNA and protein synthesis of G1 cells treated with inducer was 44-48% of that of randomly proliferating induced cells, in contrast to thymidine incorporation into DNA which decreased to 2% of that of cells replicating at the permissive temperature. Furthermore, G1 ts cells treated with inducer accumulated hemoglobin when released from growth arrest at the permissive temperature in the presence (but not in the absence) of inducer. These results are consistent with a requirement for a cell cycle-dependent event prior to commitment to differentiation.

Animals↗