Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A full-length human c-myb cDNA clone has been isolated from a CCRF-CEM leukemia cell cDNA library. The plasmid vector contains simian virus 40-derived promotor, splice, and polyadenylation sequences as well as a transcription unit for a dihydrofolate reductase cDNA. We have introduced this construct into Friend erythroleukemia (F-MEL) cells and have isolated a number of clones which contain intact and transcriptionally active human c-myb sequences. F-MEL clones expressing the highest levels of the human c-myb mRNA differentiate poorly in response to dimethyl sulfoxide. Two clones which initially expressed low levels of human c-myb transcripts and which differentiated normally were subsequently inhibited in their ability to differentiate when grown in successively higher concentrations of methotrexate, due to amplification and enhanced expression of plasmid sequences. The inhibitory effect on F-MEL differentiation appeared to be independent of the early decline in c-myc transcripts which were normally regulated in all cases examined. Our results indicate that constitutive expression of a nontruncated human c-myb cDNA can exert profound effects on erythroid differentiation and argue for a causal role of c-myb in the F-MEL differentiation process.
Mol Cell Biol 1988 Feb
PMID:Constitutive expression of a c-myb cDNA blocks Friend murine erythroleukemia cell differentiation. 283 42

Studies were carried out to examine the effect of glucocorticoids on human calpactin II (lipocortin I) mRNA expression. A cRNA probe for human calpactin II (hCPII) was used in a solution hybridization assay to study the effect of dexamethasone on hCPII mRNA levels in human skin fibroblasts, peripheral lymphocytes, pulmonary alveolar macrophages, and HeLa S3 cells. As a positive control, human metallothionein II (hMTII) mRNA levels were measured since hMTII is known to be regulated by glucocorticoids and heavy metals, both of which induce transcriptional activity of the gene. Dexamethasone treatment of these human cell types caused a dose-dependent increase in hMTII mRNA levels, whereas no effect on hCPII mRNA levels was observed. These findings were confirmed in time course studies, where 10(-6) M dexamethasone treatment caused a maximal 2- to 5-fold increase in hMTII mRNA levels after 6-8 h of treatment but no increase in hCPII mRNA levels was observed at any time point up to 24 h. A human glucocorticoid sensitive lymphoid cell line, CEM C7, and a glucocorticoid resistant mutant, ICR-27, isolated from CEM C7, were included in order to confirm the requirement of a functional glucocorticoid receptor (GR) in the induction of glucocorticoid-regulated genes. Dexamethasone (10(-6) M) induced hMTII but not hCPII mRNA in CEM C7 cells, whereas neither hMTII nor hCPII mRNA was induced in ICR-27 cells. In conclusion, our data suggest that glucocorticoids do not induce calpactin II (lipocortin I) mRNA in the human cell types studied.
Mol Endocrinol 1988 Aug
PMID:Human calpactin II (lipocortin I) messenger ribonucleic acid is not induced by glucocorticoids. 297 26

We have compared the sites of nucleotide incision on DNA damaged by oxidizing agents when cleavage is mediated by either Escherichia coli endonuclease III or an endonuclease present in bovine and human cells. E. coli endonuclease III, the bovine endonuclease isolated from calf thymus, and the human endonuclease partially purified from HeLa and CEM-C1 lymphoblastoid cells incised DNA damaged with osmium tetroxide, ionizing radiation, or high doses of UV light at sites of pyrimidines. For each damaging agent studied, regardless of whether the E. coli, bovine, or human endonuclease was used, the same sequence specificity of cleavage was observed. We detected this endonuclease activity in a variety of human fibroblasts derived from normal individuals as well as individuals with the DNA repair deficiency diseases ataxia telangiectasia and xeroderma pigmentosum. The highly conserved nature of such a DNA damage-specific endonuclease suggests that a common pathway exists in bacteria, humans, and other mammals for the reversal of certain types of oxidative DNA damage.
Mol Cell Biol 1987 Jan
PMID:A highly conserved endonuclease activity present in Escherichia coli, bovine, and human cells recognizes oxidative DNA damage at sites of pyrimidines. 303 65

The human ets-2 gene is a homolog of the v-ets oncogene of the E26 virus and codes for a 56-kilodalton nuclear protein. The ets-2 protein is phosphorylated and has a rapid turnover, with a half-life of 20 min. When human lymphocytic CEM cells were treated with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), the level of the ets-2 protein was quickly elevated 5- to 20-fold. This effect of TPA was mimicked by a synthetic diacylglycerol, 1-oleoyl-2-acetyl glycerol, and was blocked by the protein kinase C inhibitor H7, indicating that protein kinase C is involved in the induction. The increase in the ets-2 protein was due to stabilization of the protein, because the protein had a half-life of more than 2 h in the presence of TPA and the ets-2 mRNA level did not increase significantly upon TPA treatment. The protein synthesis inhibitor cycloheximide enhanced the effect of TPA on the ets-2 protein and could itself slow turnover of the protein. Properties of the ets-2 protein, such as nuclear localization, phosphorylation, rapid turnover, and response to protein kinase C, indicate that this protein belongs to a group of oncogene proteins which are generally thought to have regulatory functions in the nucleus (e.g., myc, fos, myb, and p53). Our results suggest that protein kinase C, either directly or indirectly, regulates the level of the ets-2 protein by posttranslational mechanisms.
Mol Cell Biol 1988 Nov
PMID:A short-lived nuclear phosphoprotein encoded by the human ets-2 proto-oncogene is stabilized by activation of protein kinase C. 306 67

A recombinant amphotropic retrovirus was used to introduce the protein-coding region of the IL-2 receptor cDNA derived from HUT-102 cells into human CEM leukemic T-cells that lack these receptors. CEM T-cells that contained the virus expressed functional IL-2 receptors that transiently mediated five- to tenfold increases in [3H]thymidine incorporation following the addition of picomolar quantities of IL-2. Although IL-2 responsiveness was subsequently lost, it could be reinduced by cellular activation with the OKT11 monoclonal antibody. This phenotype also proved unstable with progressive time in culture. Despite the loss of IL-2 responsiveness, the infected CEM T-cells continued to express Tac antigen and displayed 50 to 200 high-affinity IL-2 receptors per cell that bound IL-2 with a dissociation constant of 4.3 pM. This affinity is fully equivalent to that detected on activated normal T-cells (2 to 50 pM). The apparent molecular size of the Tac antigen on these cells (55,000 to 60,000 daltons) was comparable to that on normal activated T-cells but 5000 daltons larger than the aberrant IL-2 receptors on HUT-102 cells. These data demonstrate that expression of a human IL-2 receptor cDNA in human T-cells results in high-affinity IL-2 receptor display that transiently imparts an IL-2 responsive state of growth. These results also raise the possibility that the T11 surface receptor may play an important regulatory role in high-affinity IL-2 receptor expression.
Mol Biol Med 1987 Apr
PMID:Reconstitution of high affinity IL-2 receptor expression in a human T-cell line using a retroviral cDNA expression vector. 311 93

Multidrug resistance (MDR), typified by resistance to Vinca alkaloids and anthracyclines, is a well characterized experimental phenomenon that may have some clinical correlates. Verapamil, chloroquine, and related drugs have been shown previously to be capable of enhancing anticancer drug cytotoxicity in multi-drug-resistant cells, but the mechanism(s) by which these agents do this is(are) unclear. Since these agents did not seem to have common features, we studied these and other compounds for their ability to "modulate" Vinca alkaloid resistance in order to determine whether they possessed any common chemical or physical features. In addition to verapamil, 24 compounds, consisting of indole alkaloids, lysosomotropic agents, and amines, were tested for their ability to enhance the cytotoxicity of vinblastine and/or vincristine in our human leukemic multidrug-resistant cell line, CEM/VLB100. Seventeen compounds that enhance the cytotoxicity of the Vinca alkaloids by more than 5-fold have been identified. These include quinolines (chloroquine, quinine, chinchonidine, and primaquine), acridines (acridine, acridine orange, and quinacrine), and indole alkaloids (yohimbine, corynanthine, reserpine, physostigmine, and the vindoline and catharanthine moieties of the Vinca alkaloids), as well as other alkaloids and amines (chlorpromazine, propranolol, atropine, and tryptamine). Vindoline, catharanthine, and quinacrine also enhanced the cytotoxicity of doxorubicin and teniposide in these cells, indicating that this "modulation" was not limited to Vinca alkaloids. We examined some well known lysosomotropic compounds (methylamine, epinephrine, suramin, and trypan blue) and found that they were not able to enhance the cytotoxicity of vincristine in the CEM/VLB100 cells, indicating that lysosomotropic activity per se is not required for modulator activity. Three-dimensional computer modeling permitted molecular comparisons of conformationally related congeners of vinblastine, vindoline, and verapamil and revealed three regions of structural homology. We measured the hydrophobicity (by oil/water partitioning) and calculated the molar refractivity (by the additive substituent constant method) of active and inactive compounds. We found that those cationic agents--verapamil, quinacrine, indole alkaloids, and quinolines--that were lipid soluble at physiologic pH and had similar molar refractivities were best able to enhance the cytotoxicity of the Vinca alkaloids in our multidrug-resistant cells.(ABSTRACT TRUNCATED AT 400 WORDS)
Mol Pharmacol 1988 Apr
PMID:Physical-chemical properties shared by compounds that modulate multidrug resistance in human leukemic cells. 316 58

Simple and sensitive methods to directly detect the human immunodeficiency virus (HIV) are needed for routine use in the clinical laboratory. In this study, we compared DNA probes prepared by: (1) nick translation with biotinylated dATP; (2) direct covalent biotinylation with photobiotin; (3) direct covalent reaction with 2-acetylaminofluorene (AAF); and (4) a standard radioactive (32P) nick translation procedure. These four DNA probes were hybridized with dilutions of purified target HIV DNA blotted onto nitrocellulose strips. Hybridization was detected using a complex of strepavidin-alkaline phosphatase [for (1) and (2)], alkaline phosphatase-tagged antibodies [for (3)] and by autoradiography [for (4)]. Alkaline phosphatase was detected colorimetrically using nitroblue tetrazolium and 5-bromo-4-chloro-3-indolyl phosphate. After 1 h, AAF probes were most sensitive (amount detected less than 5 pg), followed by biotin (10 pg), photobiotinylated probes (20 pg) and the radioactive probe (10 pg). The AAF probes were then used to detect HIV DNA in infected CEM cells. We conclude that non-radioactive DNA labelling methods can be used to directly detect HIV DNA under conditions compatible with present clinical laboratory procedures.
Mol Cell Probes 1987 Dec
PMID:Comparison of non-radioactive DNA hybridization probes to detect human immunodeficiency virus nucleic acid. 345 25

Somatic hybrids between two glucocorticoid-resistant clonal cell lines, CEM C1 and ICR-27, isolated independently from the CCRF-CEM human lymphoblastoid cell line, were constructed to investigate the complementation effect between the glucocorticoid receptor and the gene product(s) for inducing receptor-mediated lymphocytolysis. CEM C1 (r+ly-) has a normal amount of functional glucocorticoid receptor as compared to the steroid-sensitive clonal line CEM C7. Clone ICR-27 (r-ly?), which was originally isolated following mutagenesis of CEM C7 with the mutagen ICR 191, has few glucocorticoid receptors as determined by whole-cell receptor assay. The eight randomly selected CEM C1 X ICR-27 hybrid clones all showed sensitivity to 10(-6) M dexamethasone (ly+). The receptor site content of two near-tetraploid hybrids chosen for analysis was close to that of CEM C1 (r+). Hybrids constructed between CEM C1 and the receptor-bearing, steroid-sensitive clone, CEM C7 (r+ly+) also showed glucocorticoid sensitivity, and their receptor sites corresponded to the sum of those of CEM C1 and CEM C7 (r+r+ly+). These results indicate: that CEM C1 has no trans-active inhibitor of lysis; that CEM C1 has intact glucocorticoid receptor; and that ICR-27 and CEM C1 complement one another to restore lymphocytolysis. Therefore, CEM C1 cells can serve as a donor of human glucocorticoid receptors.
Somat Cell Mol Genet 1987 Jan
PMID:Complementation between glucocorticoid receptor and lymphocytolysis in somatic cell hybrids of two glucocorticoid-resistant human leukemic clonal cell lines. 346 32

The metabolic consequences of deoxyuridine treatment in four cultured human lymphoblast lines (CCRF-CEM, RPMI-8402, JM, and BALM) were studied by cell growth experiments, flow cytometry, and measurement of 2'-deoxyribonucleoside triphosphate (dNTP) levels. DNA perturbations occurred in all lymphoblast lines, but there was no significant impairment of RNA synthesis. The DNA perturbations in CCRF-CEM, RPMI-8402, and BALM cells reflected inhibition of DNA synthesis, and the associated dNTP changes were consistent with ribonucleotide reductase inhibition or, specifically in BALM cells, with DNA alpha-polymerase inhibition. JM cells treated with an intermediate concentration of deoxyuridine developed a block at the G1/S boundary which was deoxyuridine concentration-dependent, but not specific for deoxyuridine (it was also seen with thymidine treatment) and not related to DNA synthesis inhibition. There were no idiosyncratic dNTP effects accompanying the G1/S boundary block, and the responsible metabolic mechanism remains to be determined.
Mol Pharmacol 1982 Jul
PMID:The metabolic basis of deoxyuridine cytotoxicity. Studies of cultured human lymphoblasts. 618 85

A cDNA cloning approach was used to study the regulation of gene expression in human T lymphocytes upon mitogen stimulation. Poly(A)+ mRNA was prepared from phytohemagglutinin A (PHA) and 12-O-tetradecanoyl phorbol 13-acetate (TPA) activated human T cells and a cDNA clone library was constructed. After screening by colony hybridization with [32P]cDNA probes made from resting and activated T cell mRNA, several clones whose mRNA increased at least 10 to 20-fold upon stimulation were isolated. Northern blot analysis of the mRNA from various cell types using these cDNA clones as probes revealed that one of the cDNA clones, pNC5A, encoded a gene expressed only in PHA and TPA-stimulated human T lymphocytes and in a human neoplastic T cell line HUT102-SH9. Less than 20 copies of this mRNA species per cell was detected in resting human T lymphocytes, B lymphocytes and monocytes and in two other T cell lymphoma lines (CEM and MOLT4), two B lymphoblastoid cell lines (WIL2-729-HF2 and HFB-1), a myeloid cell line (HL60) and a human embryonic lung fibroblast cell line (MRC-5). Hybrid selection translation and sodium dodecyl sulfate/polyacrylamide gel electrophoretic analysis of the translated product indicated that a polypeptide of 30,000 to 32,000 Mr is encoded by this particular cDNA clone. Thus, this cDNA clone may define a novel gene that is expressed only in activated human T cells.
Mol Biol Med 1984 Apr
PMID:A cDNA clone encoding a product of activated human T lymphocytes. 633 9


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