Glucose-regulated protein-78 expression in the rat adrenal cortex

1993 ◽  
Vol 10 (1) ◽  
pp. 33-42 ◽  
Author(s):  
J Gregoire ◽  
L M Mertz ◽  
R C Pedersen
Keyword(s):  
Steady State ◽  
Sequence Similarity ◽  
Cdna Probe ◽  
The Other ◽  
Northern Analysis ◽  
Intracellular Accumulation ◽  
Adrenocortical Cells ◽  
Steady State Levels ◽  
Carboxyl Terminal ◽  
Glucose Regulated Protein

ABSTRACT We have postulated that steroidogenesis activator polypeptide (SAP) is a product of glucose-regulated protein-78 (grp78) proteolysis on the basis of a number of considerations, including a striking sequence similarity between the carboxyl-terminal region of grp78 and SAP. Since ACTH stimulates the rapid intracellular accumulation of SAP, experiments were conducted to determine whether ACTH might also regulate levels of grp78 mRNA and/or protein. Using a grp78 cDNA probe, Northern analysis of total RNA isolated from hypophysectomized or dexamethasone-suppressed rats revealed that neither treatment had a measurable influence on steady-state levels of grp78 mRNA over a 4-day period. Moreover, immunoblotting with an antiserum directed against a shared grp78/SAP sequence failed to detect a significant change in the grp78 content of adrenal homogenates from dexamethasone-suppressed rats as compared with untreated controls. On the other hand, grp78 in cultured rat adrenocortical cells fell to 50% of that in time-zero controls after 72 h in the absence of ACTH, whereas inclusion of ACTH in the medium blocked this decline. We conclude that while adrenocortical grp78 may be under some measure of trophic ACTH control, the rapid fluctuations reported for SAP are not likely to be driven by large changes in the size of the grp78 pool.

scholarly journals Cloning and characterization of a complementary DNA for a thyroid hormone-responsive protein in mature rat cerebral tissue

1997 ◽  
Vol 327 (2) ◽  
pp. 617-623 ◽  
Author(s):  
N. Gul SHAH ◽  
Jianping LI ◽  
Patricia SCHNEIDERJOHN ◽  
D. Arshag MOORADIAN
Keyword(s):  
Thyroid Hormone ◽  
Rat Brain ◽  
Molecular Mechanisms ◽  
Sequence Similarity ◽  
Cdna Probe ◽  
Northern Analysis ◽  
Adrenergic System ◽  
Rat Tissues ◽  
Cerebral Tissue ◽  
Western Analysis

A gene responsive to thyroid hormone (TH) has been identified in the adult rat brain cerebral tissue. A cDNA probe differentially expressed in euthyroid, hypothyroid and hyperthyroid rat cerebral tissue, generated by reverse transcriptase-PCR differential display of mRNA, was used to screen the rat brain cDNA library. A 3.4 kb positive clone hybridized in Northern blots with a 3.8 kb mRNA that proved to be TH responsive (THR). The remaining coding sequence and a part of the 5ʹ untranslated region of this cDNA were obtained by 5ʹ rapid amplification of cDNA ends. The deduced amino acid sequence revealed that THR protein (THRP), a 68 kDa moiety, has 83% sequence similarity with c-Abl interactor protein (Abi-2), which is a substrate for tyrosine kinase activity of c-Abl. The extensive similarity between the two proteins suggests a potential role for THRP as a substrate for c-Abl. Northern analysis showed that the expression of THR mRNA in hyperthyroid rats is 6-fold that in euthyroid rats. There is also a 4-6-fold increase in the concentration of THRP, as analysed by Western analysis. Owing to the extensive similarity between rat THRP and human Abi-2, a polyclonal anti- (human Abi-2) antibody was successfully used for Western analysis of proteins from the rat tissues. The observed increase in both the mRNA and the protein did not decline after β-adrenergic system blockade with propranolol, suggesting that the action of TH on the expression of this gene is not mediated through the β-adrenergic system. Immunohistochemical studies revealed that neuronal cells were particularly rich in THRP. Both THR mRNA and THRP are rapidly induced in vivo after intravenous administration of thyroxine. Tissue distribution studies indicated that the cerebral tissue was particularly enriched with THR mRNA and 68 kDa THRP. A cDNA clone for a THR gene could provide a useful tool to study the molecular mechanisms of TH effects on cerebral tissue in adult animals.

Drosophila melanogaster homologs of the raf oncogene

1987 ◽  
Vol 7 (6) ◽  
pp. 2134-2140
Author(s):  
G E Mark ◽  
R J MacIntyre ◽  
M E Digan ◽  
L Ambrosio ◽  
N Perrimon
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Steady State ◽  
Kinase Domain ◽  
Early Embryogenesis ◽  
The Other ◽  
Chromosome 2 ◽  
Related Gene ◽  
Steady State Levels

A murine v-raf probe, representing the kinase domain, was used to identify two unique loci in Drosophila melanogaster DNA. The most closely related to v-raf was mapped by in situ hybridization to position 2F5-6 (Draf-1) on the X chromosome, whereas the other raf-related gene (Draf-2) was found at position 43A2-5 on chromosome 2. The nucleotide and amino acid homologies of Draf-1 to the kinase domain of v-raf are 61 and 65%, respectively. The large amount of a 3.2-kilobase Draf-1 transcript detected in eggs as a maternal message decreases during embryonic development, and significant steady-state levels are observed throughout the remainder of morphogenesis. We speculate that the Draf-1 locus plays an important role in early embryogenesis.

The DAL81 gene product is required for induced expression of two differently regulated nitrogen catabolic genes in Saccharomyces cerevisiae

1991 ◽  
Vol 11 (2) ◽  
pp. 1161-1166 ◽  
Author(s):  
P A Bricmont ◽  
J R Daugherty ◽  
T G Cooper
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Steady State ◽  
Amino Acid ◽  
The Other ◽  
Loss Of Function ◽  
Induced Expression ◽  
Catabolic Genes ◽  
Acid Protein ◽  
Steady State Levels ◽  
Amino Acid Protein

We demonstrate that the DAL81 gene, previously thought to be specifically required for induced expression of the allantoin pathway genes in Saccharomyces cerevisiae, functions in a more global manner. The data presented show it to be required for utilization of 4-aminobutyrate as a nitrogen source and for 4-aminobutyrate-induced increases in the steady-state levels of UGA1 mRNA. The DAL81 gene encodes a 970-amino-acid protein containing sequences homologous to the Zn(II)2Cys6 motif and two stretches of polyglutamine residues. Deletion of sequences homologous to the Zn(II)2Cys6 motif did not result in a detectable loss of function. On the other hand, loss of one of the polyglutamine stretches, but not the other, resulted in a 50% loss of DAL81 function.

scholarly journals The DAL81 gene product is required for induced expression of two differently regulated nitrogen catabolic genes in Saccharomyces cerevisiae.

1991 ◽  
Vol 11 (2) ◽  
pp. 1161-1166 ◽  
Author(s):  
P A Bricmont ◽  
J R Daugherty ◽  
T G Cooper
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Steady State ◽  
Amino Acid ◽  
The Other ◽  
Loss Of Function ◽  
Induced Expression ◽  
Catabolic Genes ◽  
Acid Protein ◽  
Steady State Levels ◽  
Amino Acid Protein

We demonstrate that the DAL81 gene, previously thought to be specifically required for induced expression of the allantoin pathway genes in Saccharomyces cerevisiae, functions in a more global manner. The data presented show it to be required for utilization of 4-aminobutyrate as a nitrogen source and for 4-aminobutyrate-induced increases in the steady-state levels of UGA1 mRNA. The DAL81 gene encodes a 970-amino-acid protein containing sequences homologous to the Zn(II)2Cys6 motif and two stretches of polyglutamine residues. Deletion of sequences homologous to the Zn(II)2Cys6 motif did not result in a detectable loss of function. On the other hand, loss of one of the polyglutamine stretches, but not the other, resulted in a 50% loss of DAL81 function.

scholarly journals Hormonal regulation of collagenolysis in uterine cervical fibroblasts. Modulation of synthesis of procollagenase, prostromelysin and tissue inhibitor of metalloproteinases (TIMP) by progesterone and oestradiol-17 β

1991 ◽  
Vol 275 (3) ◽  
pp. 645-650 ◽  
Author(s):  
T Sato ◽  
A Ito ◽  
Y Mori ◽  
K Yamashita ◽  
T Hayakawa ◽  
...  
Keyword(s):  
Steady State ◽  
Hormonal Regulation ◽  
Culture Media ◽  
Interleukin 1 ◽  
State Level ◽  
The Other ◽  
Steady State Level ◽  
Tissue Inhibitor Of Metalloproteinases ◽  
Tissue Inhibitor ◽  
Steady State Levels

Rabbit uterine cervical fibroblasts produced a large amount of matrix metalloproteinases (MMPs) such as collagenase (MMP-1) and stromelysin (MMP-3) and a small relatively amount of tissue inhibitor of metalloproteinases (TIMP). When cells were treated with progesterone or oestradiol-17 beta, both steroids concurrently decreased the level of procollagenase and prostromelysin in the culture media and the steady-state levels of the respective mRNAs. On the other hand, the level of TIMP in the culture media and the steady-state level of its mRNA were simultaneously increased by these steroids. Similarly, the suppression of production of MMPs and the augmentation of TIMP production by both steroids were observed with interleukin 1 (IL-1)-treated cells, but the action of progesterone was more effective than that of oestradiol-17 beta in the IL-1-untreated and -treated cells. These results suggest that collagenolysis in uterine cervical fibroblasts is negatively regulated by steroid hormones via the acceleration of TIMP production and the suppression of synthesis of MMPs at the pretranslational level.

scholarly journals Mouse Adenovirus Type 1 Early Region 1A Is Dispensable for Growth in Cultured Fibroblasts

1998 ◽  
Vol 72 (8) ◽  
pp. 6325-6331 ◽  
Author(s):  
Baoling Ying ◽  
Kimberley Smith ◽  
Katherine R. Spindler
Keyword(s):  
Steady State ◽  
Adenovirus Type ◽  
Wild Type Virus ◽  
Northern Analysis ◽  
Mrna Levels ◽  
Wild Type ◽  
Cultured Fibroblasts ◽  
Early Region ◽  
Steady State Levels

ABSTRACT Mouse adenovirus type 1 (MAV-1) mutants with deletions of conserved regions of early region 1A (E1A) or with point mutations that eliminate translation of E1A were used to determine the role of E1A in MAV-1 replication. MAV-1 E1A mutants expressing no E1A protein grew to titers comparable to wild-type MAV-1 titers on mouse fibroblasts (3T6 fibroblasts and fibroblasts derived from Rb+/+,Rb+/−, and Rb−/− transgenic embryos). To test the hypothesis that E1A could induce a quiescent cell to reenter the cell cycle, fibroblasts were serum starved to stop DNA replication and cellular replication and then infected with the E1A mutant and wild-type viruses. All grew to equivalent titers. Steady-state levels of MAV-1 early mRNAs (E1A, E1B, E2, E3, and E4) from 3T6 cells infected with wild-type or E1A mutant virus were examined by Northern analysis. Steady-state levels of mRNAs from the mutant-infected cells were comparable to or greater than the levels found in wild-type virus infections for most of the early regions and for two late genes. The E2 mRNA levels were slightly reduced in all mutant infections relative to wild-type infections. E1A mRNA was not detected from infections with the MAV-1 E1A null mutant, pmE109, or from infections with similar MAV-1 E1A null mutants, pmE112 andpmE113. The implications for the lack of a requirement of E1A in cell culture are discussed.

scholarly journals The Dynamic Free Rider Problem: A Laboratory Study

2016 ◽  
Vol 8 (4) ◽  
pp. 268-308 ◽  
Author(s):  
Marco Battaglini ◽  
Salvatore Nunnari ◽  
Thomas R. Palfrey
Keyword(s):  
Steady State ◽  
Public Goods ◽  
Public Good ◽  
Free Rider ◽  
The Other ◽  
The Public ◽  
Free Rider Problem ◽  
Markov Perfect ◽  
Steady State Levels ◽  
Perfect Equilibria

We report the results of an experiment that investigates free riding in the accumulation of durable public goods. We consider economies with reversibility, where contributions can be positive or negative; and economies with irreversibility, where contributions are nonnegative. Aggregate outcomes support the qualitative predictions of the Markov Perfect Equilibria (MPE) characterized in Battaglini, Nunnari, and Palfrey (2014): steady state levels of public good are lower with reversibility than irreversibility; accumulation is inefficiently slow; and the public good is under-provided in both regimes. On the other hand, public good levels are higher than MPE, and some evidence of history dependence is detected. (JEL C91, H41)

scholarly journals Characterization of the Proteasome Accessory Factor (paf) Operon in Mycobacterium tuberculosis

2007 ◽  
Vol 189 (8) ◽  
pp. 3044-3050 ◽  
Author(s):  
Richard A. Festa ◽  
Michael J. Pearce ◽  
K. Heran Darwin
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Steady State ◽  
Unknown Function ◽  
The Other ◽  
Accessory Factor ◽  
Reactive Nitrogen Intermediates ◽  
Steady State Levels ◽  
The Stability ◽  
Transposon Insertions

ABSTRACT In a previous screen for Mycobacterium tuberculosis mutants that are hypersusceptible to reactive nitrogen intermediates (RNI), two genes associated with the M. tuberculosis proteasome were identified. One of these genes, pafA (proteasome accessory factor A), encodes a protein of unknown function. In this work, we determined that pafA is in an operon with two additional genes, pafB and pafC. In order to assess the contribution of these genes to RNI resistance, we isolated mutants with transposon insertions in pafB and pafC. In contrast to the pafA mutant, the pafB and pafC mutants were not severely sensitized to RNI, but pafB and pafC were nonetheless required for full RNI resistance. We also found that PafB and PafC interact with each other and that each is likely required for the stability of the other protein in M. tuberculosis. Finally, we show that the presence of PafA, but not PafB or PafC, regulates the steady-state levels of three proteasome substrates. Taken together, these data demonstrate that PafA, but not PafB or PafC, is critical for maintaining the steady-state levels of known proteasome substrates, whereas all three proteins appear to play a role in RNI resistance.

scholarly journals Stimulation of Osteoprotegerin Ligand and Inhibition of Osteoprotegerin Production by Glucocorticoids in Human Osteoblastic Lineage Cells: Potential Paracrine Mechanisms of Glucocorticoid-Induced Osteoporosis1

Endocrinology ◽  
1999 ◽  
Vol 140 (10) ◽  
pp. 4382-4389 ◽  
Author(s):  
Lorenz C. Hofbauer ◽  
Francesca Gori ◽  
B. Lawrence Riggs ◽  
David L. Lacey ◽  
Colin R. Dunstan ◽  
...  
Keyword(s):  
Steady State ◽  
Bone Resorption ◽  
Conditioned Medium ◽  
Northern Analysis ◽  
Protein Synthesis Inhibitor ◽  
Mrna Levels ◽  
Trap Activity ◽  
Osteoblastic Lineage ◽  
Steady State Levels

Abstract Osteoporosis is a serious complication of systemic glucocorticoid use. However, while glucocorticoids increase bone resorption in vitro and in vivo, the mechanism(s) of this effect are at present unclear. Recent studies have identified the osteoprotegerin (OPG) ligand (OPG-L) as the final effector of osteoclastogenesis, an action that is opposed by the soluble neutralizing receptor, OPG. Thus, we assessed glucocorticoid regulation of OPG and OPG-L in various human osteoblastic lineage cells using Northern analysis, RT-PCR, and ELISA. Dexamethasone inhibited constitutive OPG messenger RNA (mRNA) steady-state levels by 70–90% in primary (MS) and immortalized stromal cells (hMS), primary trabecular osteoblasts (hOB), immortalized fetal osteoblasts (hFOB), and osteosarcoma cells (MG-63). In hFOB cells, dexamethasone inhibited constitutive OPG mRNA steady-state levels in a dose- and time-dependent fashion by 90%, and also suppressed cytokine-stimulated OPG mRNA steady-state levels. Dexamethasone-induced inhibition of OPG mRNA levels was not affected by the protein synthesis inhibitor, cycloheximide, and was shown to be due to inhibition of OPG gene transcription using a nuclear run-on assay. Moreover, dexamethasone also dose dependently (10−10m–10−7m) inhibited constitutive OPG protein concentrations in the conditioned medium of hFOB cells from 2.59 ± 0.02 ng/ml (control) to 0.30 ± 0.01 ng/ml (88% inhibition; P < 0.001 by ANOVA). Concurrently, dexamethasone stimulated OPG-L mRNA steady-state levels in MS and hFOB cells by 2- and 4-fold, respectively. Treatment of murine marrow cultures with conditioned medium harvested from dexamethasone-treated MG-63 cells increased tartrate-resistant acid phosphatase (TRAP) activity by 54% (P < 0.005) compared with medium harvested from control-treated cells (in the presence of OPG-L and macrophage colony-stimulating factor). Moreover, dexamethasone (10−8m) promoted osteoclast formation in vitro, as assessed by a 2.5-fold increase of TRAP activity in cell lysates (P < 0.001) and the appearance of TRAP-positive multinucleated cells. Our data are thus consistent with the hypothesis that glucocorticoids promote osteoclastogenesis by inhibiting OPG and concurrently stimulating OPG-L production by osteoblastic lineage cells, thereby enhancing bone resorption.

scholarly journals Drosophila melanogaster homologs of the raf oncogene.

1987 ◽  
Vol 7 (6) ◽  
pp. 2134-2140 ◽  
Author(s):  
G E Mark ◽  
R J MacIntyre ◽  
M E Digan ◽  
L Ambrosio ◽  
N Perrimon
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Steady State ◽  
Kinase Domain ◽  
Early Embryogenesis ◽  
The Other ◽  
Chromosome 2 ◽  
Related Gene ◽  
Steady State Levels

A murine v-raf probe, representing the kinase domain, was used to identify two unique loci in Drosophila melanogaster DNA. The most closely related to v-raf was mapped by in situ hybridization to position 2F5-6 (Draf-1) on the X chromosome, whereas the other raf-related gene (Draf-2) was found at position 43A2-5 on chromosome 2. The nucleotide and amino acid homologies of Draf-1 to the kinase domain of v-raf are 61 and 65%, respectively. The large amount of a 3.2-kilobase Draf-1 transcript detected in eggs as a maternal message decreases during embryonic development, and significant steady-state levels are observed throughout the remainder of morphogenesis. We speculate that the Draf-1 locus plays an important role in early embryogenesis.

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