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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Picornavirus infection induces a profound inhibition of labelling of newly synthesized RNA in some cell lines. EMC virus blocks transcription in L929 cells, particularly at early times during infection. This inhibition is not dependent on virus gene expression, since it occurs with UV-inactivated virus and also in the presence of translation inhibitors. The inhibition can be largely accounted for by the blockade of [3H]nucleoside transport, as suggested by the transport kinetics and incorporation of labelled nucleoside from preloaded cells. The inhibition of transport and incorporation into TCA-precipitable material was observed with pyrimidine (uridine, thymidine and cytosine) and purine nucleosides (adenosine and guanosine), but the blockade by EMC virus was higher with the latter nucleosides. Preloading of cells with any of these nucleosides resulted in a decreased effect on nucleoside incorporation into nucleic acid after virus infection. These results suggest that the inhibition of incorporation of labelled nucleosides into nucleic acid in EMC virus-infected cells can be explained, at least in part, by the decreased pool size of the phosphorylated nucleosides. These effects are not specific for L cells, because they are also observed in other cell lines.
Mol Cell Biochem 1986 Jun
PMID:The inhibition of nucleic acid synthesis in encephalomyocarditis virus-infected L929 cells: effects on nucleoside transport. 242 45

Pig thyroid slices, pre-labelled with [125I]iodide, were incubated with or without thyrotropin (TSH), dibutyryl cyclic AMP (dbcAMP) or forskolin. After lysosome isolation, intralysosomal thyroglobulin (Tg) hydrolysis was determined by the increment in trichloroacetic acid (TCA)-soluble radioactivity. TSH stimulated the intralysosomal Tg hydrolysis. This stimulation was time and concentration dependent and was mimicked by forskolin or dbcAMP. When endocytosis and protein synthesis were blocked by inhibitors (nocodazole and puromycin) the stimulatory effect was still maintained. We conclude that TSH increases quickly and specifically, via a cAMP-mediated process, intralysosomal Tg hydrolysis, independent of its effects on endocytosis of Tg and lysosomal protease synthesis.
Mol Cell Endocrinol 1987 Jan
PMID:Intralysosomal hydrolysis of thyroglobulin: specific and cAMP-mediated activation by TSH. 243 87

Exposure of human decidual cells for 0.5 h to dibutyryl cAMP, isobutyl-methylxanthine (IBMX), cholera toxin or forskolin caused a dose-dependent inhibition of prolactin release with maximal inhibition by each agent of 50-60%. Dibutyryl cAMP (5 mM), IBMX (0.5 mM), and cholera toxin (10 micrograms/ml) also inhibited prolactin synthesis to the same extent as prolactin release. Dibutyryl cAMP, IBMX, and cholera toxin, however, had no effect on the release of 35S-methionyl-prolactin from decidual cells preincubated for 24 h in medium with 35S-methionine. These agents, however, had no effects on the synthesis or release of TCA-precipitable 35S-decidual proteins and did not cause the degradation of intracellular or released prolactin. The demonstration that agents which increase intracellular cAMP levels inhibit the synthesis and release of decidual prolactin strongly implicates cAMP as a second messenger in the regulation of the synthesis and release of the hormone. The inhibitory effect of cAMP on prolactin release appears to be on the release from a rapidly releasable, newly synthesized intracellular prolactin pool.
Mol Cell Endocrinol 1987 Mar
PMID:Cyclic AMP inhibits the synthesis and release of prolactin from human decidual cells. 243 70

We isolated a cDNA clone from Arabidopsis thaliana encoding the TCA cycle enzyme, citrate synthase. The plant enzyme displays 48% and 44% amino acid residue similarity with the pig, and yeast polypeptides, respectively. Many proteins, including citrate synthase, which are destined to reside in organelles such as mitochondria and chloroplasts, are the products of the nucleocytoplasmic protein synthesizing machinery and are imported post-translationally to the site of function. We present preliminary investigations toward the establishment of an in vitro plant mitochondrial import system allowing for future studies to dissect this process in plants where the cell must differentiate between mitochondria and chloroplast and direct their polypeptides appropriately.
Plant Mol Biol 1989 Oct
PMID:Isolation of a cDNA encoding mitochondrial citrate synthase from Arabidopsis thaliana. 249 64

The products released by Leishmania major promastigotes incubated with [1-13C]glucose as sole exogenous carbon source were identified using nuclear magnetic resonance (NMR). Under aerobic (95% O2/5% CO2) conditions, acetate, succinate, and small amounts of pyruvate, D-lactate, and glycerol were released in addition to CO2. Under anaerobic (95% N2/5% CO2) conditions, the relative amounts of products formed changed and alanine was also released. The changes in the rates of glucose consumption and product formation during the aerobic to anaerobic transition were measured. Under hypoxic conditions (O2 less than 0.2%), glucose consumption was decreased by about 50%. Under completely anaerobic conditions (100% N2), glucose consumption almost ceased (a total reverse Pasteur effect). The inclusion of 5% CO2 in the gas phase restored hypoxic and anaerobic glucose consumption to the aerobic rate, and increased production of succinate, pyruvate, and D-lactate. Thus, CO2 and very low concentrations of O2 have strong regulatory effects on L. major glucose metabolism. A quantitative carbon balance showed that the NMR-identified products accounted for only about 25% of the glucose carbons consumed under aerobic conditions. CO2, measured as the release of 14CO2 from [U-14C]glucose, accounted for an additional 25% of the glucose consumed. About 11% of the glucose carbon was incorporated into trichloroacetic acid-insoluble products, mostly lipid. Large amounts of label from [U-14C]glucose were incorporated into the intracellular pools of alanine, glutamate, glutamine, and aspartate, indicating that CO2 from unlabeled amino acids contributed to the carbon balance. Under anaerobic conditions, all the glucose carbons consumed could be accounted for solely by the NMR-identified products.
Mol Biochem Parasitol 1989 Mar 01
PMID:Carbon dioxide abolishes the reverse Pasteur effect in Leishmania major promastigotes. 249 56

Studies were conducted to assess the role of catecholestrogens on ovarian follicular growth using cultured porcine granulosa cells. Effects of the catecholestrogens, 2-hydroxyestradiol (2-OH-E2) and 2-methoxyestradiol (2-MeO-E2) were compared to those of estradiol (E2). Treatment with saturating concentrations of 2-OH-E2 caused a significantly greater decrease in cell numbers measured after 2 days of treatment than E2 treatment. The inhibitory effect of 2-OH-E2 was time and concentration dependent, not associated with a change in the viability of cells, and was partially reversible. The potency of 2-MeO-E2 to inhibit cell numbers was similar to or greater than that of 2-OH-E2. 2-MeO-E2 had a greater inhibitory effect on DNA synthesis, as measured by [3H]thymidine incorporation into trichloroacetic acid-precipitable macromolecules, than 2-OH-E2 or E2 in the absence or presence of insulin, epidermal growth factor or platelet-derived growth factor. Concurrent treatment with epinephrine significantly enhanced the inhibitory effect of 2-OH-E2 on granulosa cell DNA synthesis. Collectively, these studies indicate that catecholestrogens are more potent inhibitors of granulosa cell replication than E2 and 5 alpha-dihydrotestosterone (DHT), and that catecholamines may modulate the antimitotic activity of 2-OH-E2. These results support the hypothesis that catecholestrogens play a role in proliferation of granulosa cells during growth of ovarian follicles.
Mol Cell Endocrinol 1989 Jun
PMID:Catecholestrogens inhibit proliferation and DNA synthesis of porcine granulosa cells in vitro: comparison with estradiol, 5 alpha-dihydrotestosterone, gonadotropins and catecholamines. 254 74

The type II pneumocyte plays a principle role in the maintenance and repair of the pulmonary alveolar epithelium by increasing its rate of proliferation under conditions of epithelial damage. This investigation examined the role of the alveolar macrophage in the control of type II cell division through its ability to produce specific growth factors when activated in vitro. Type II cells were isolated from adult male rabbits and cultured in the presence of media and matrix that support cell proliferation. Proliferation was assessed by cell counting and pulsing with [3H]thymidine, followed by measurements of labeling index and TCA-insoluble radioactivity. Alveolar macrophages were cultured in serum-free media in the presence of a particulate stimulus. Conditioned media was diluted and added to type II cell cultures. Conditioned media from stimulated macrophage cultures was found to double basal type II cell proliferation, whereas media from unstimulated macrophage cultures had no effect. Macrophage production of type II cell growth-promoting activity was dependent on the concentration of the stimulus and the length of the incubation. Investigation into the identity of the growth-regulating protein established that it is heat labile, insensitive to reduction and acidic conditions, and sensitive to trypsin digestion. Its molecular weight appears to be greater than or equal to 25 kD. Addition of several characterized growth factors to type II cell cultures demonstrated that other known growth-promoting products of macrophages do not act as type II cell growth factors. The evidence presented suggests that in vitro activated alveolar macrophages produce a type II cell growth factor that may play a critical role in mediating repair of the alveolar epithelium.
Am J Respir Cell Mol Biol 1989 Aug
PMID:Stimulated rabbit alveolar macrophages secrete a growth factor for type II pneumocytes. 261 98

We describe a new method for accurately defining the sequence recognition properties of DNA-binding proteins by selecting high-affinity binding sites from random-sequence DNA. The yeast transcriptional activator protein GCN4 was coupled to a Sepharose column, and binding sites were isolated by passing short, random-sequence oligonucleotides over the column and eluting them with increasing salt concentrations. Of 43 specifically bound oligonucleotides, 40 contained the symmetric sequence TGA(C/G)TCA, whereas the other 3 contained sequences matching six of these seven bases. The extreme preference for this 7-base-pair sequence suggests that each position directly contacts GCN4. The three nucleotide positions on each side of this core heptanucleotide also showed sequence preferences, indicating their effect on GCN4 binding. Interestingly, deviations in the core and a stronger sequence preference in the flanking region were found on one side of the central C . G base pair. Although GCN4 binds as a dimer, this asymmetry supports a model in which interactions on each side of the binding site are not equivalent. The random selection method should prove generally useful for defining the specificities of other DNA-binding proteins and for identifying putative target sequences from genomic DNA.
Mol Cell Biol 1989 Jul
PMID:Defining the sequence specificity of DNA-binding proteins by selecting binding sites from random-sequence oligonucleotides: analysis of yeast GCN4 protein. 267 75

We demonstrate the differential sensitivity of poorly differentiated and well differentiated human colon carcinoma cells to nutrients alone or to nutrients and polypeptide growth factors under completely serum-free conditions. 3H-Thymidine incorporation into trichloroacetic acid precipitable material and autoradiographic analysis indicated that nutrient replenishment alone was sufficient to initiate DNA synthesis in quiescent poorly differentiated cells, whereas defined polypeptide growth factors produced no additional effect. In contrast, well differentiated cells were mitogenically stimulated to a much greater extent by growth factors (epidermal growth factor + insulin + transferrin), than by nutrient replenishment alone. Expression of the c-myc protooncogene was increased approximately 5-fold after growth factor addition to the well differentiated cells. Maximal expression of c-myc occurred at 4 h post stimulation. In contrast, nutrients resulted in only a slight up-regulation of c-myc (1.8-fold) at approximately 90 min after addition. Addition of nutrients and/or growth factors to the poorly differentiated colon carcinoma cells resulted in an initial decline in c-myc expression (90 min), presumably due to removal of endogenous growth stimulators. Expression of c-myc returned to baseline levels by 24 h after additions. The results indicate that differential sensitivity to polypeptide growth factors is related to differentiation status in this model system and suggest that the insensitivity of poorly differentiated cells to exogenous growth factors may be due to a greater production of autocrine growth stimulators.
Mol Endocrinol 1989 Aug
PMID:Effects of growth stimulatory factors on mitogenicity and c-myc expression in poorly differentiated and well differentiated human colon carcinoma cells. 267 94

To determine whether 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] regulates transcription of the rat renal calbindin-D28k gene, the rate of calbindin-D28k mRNA synthesis was measured directly in nuclei using the in vitro nuclear transcription assay. Nuclei were prepared from kidneys of vitamin D-deficient rats at various times after a single ip injection of 1,25-(OH)2D3, and transcription was allowed to proceed in vitro in the presence of [32P]UTP for 30 min at 29 C, at which time the incorporation of UTP into trichloroacetic acid-precipitable material was optimal. Incorporation of UTP was decreased by 64.6% by alpha-amanitin, which selectively inhibits polymerase II. Purified [32P]RNA was analyzed for newly synthesized calbindin-D-28k gene transcripts by hybridization to calbindin-D28k cDNA immobilized on nitrocellulose filters. Using this assay we found that the first significant increase in calbindin-D28k gene transcription occurred at 1 h, and the peak of transcriptional activity occurred at 2 h. Within 12 h of 1,25-(OH)2D3 treatment, calbindin-D28k gene transcription returned to control levels. Using Northern blot analysis, a significant increase in calbindin-D RNA was first observed 2 h after hormone administration, reaching a maximum at 12 h. Renal calbindin-D28k protein levels are significantly increased by 3 h and reach a maximum value 48 h after hormone administration. Our results suggest that the early increase in renal calbindin-D28k may be due to transcriptional regulation. The long time lag between transcription and the peak of calbindin mRNA and calbindin protein accumulation may reflect the involvement of post-transcriptional mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS)
Mol Endocrinol 1989 Mar
PMID:Transcriptional regulation and chromosomal assignment of the mammalian calbindin-D28k gene. 274 55


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