Regulation of the mRNA for monocyte-derived neutrophil-activating peptide in differentiating HL60 promyelocytes

1989 ◽  
Vol 9 (5) ◽  
pp. 1946-1957
Author(s):  
J Kowalski ◽  
D T Denhardt
Keyword(s):  
Sodium Butyrate ◽  
Mrna Stability ◽  
Untranslated Region ◽  
Actinomycin D ◽  
Human Tumor ◽  
Lung Fibroblasts ◽  
Macrophage Differentiation ◽  
Cytoplasmic Rna ◽  
Transcriptional Induction ◽  
Full Length Clone

A cDNA library was constructed from HL60 human promyelocyte poly(A)+ RNA harvested 3 h after induction of macrophage differentiation with 12-O-tetradecanoyl phorbol-13-acetate in the presence of cycloheximide. We isolated from this library a 1.6-kilobase full-length clone designated b4 whose corresponding mRNA was greatly increased in abundance in cytoplasmic RNA under these conditions. Dideoxy sequencing revealed that this mRNA encoded MONAP (monocyte-derived neutrophil-activating peptide), a 10-kilodalton monokine with neutrophil-specific chemotactic and enzyme-releasing activities. The 3' untranslated region of this mRNA was found to be 1.2 kilobases long and possessed nine copies of the AUUUA sequence known to be associated with regulation of mRNA stability. Actinomycin D chase experiments yielded evidence that cytoplasmic stabilization was one of the means of regulation of MONAP expression. Analysis of cytoplasmic poly(A)- RNA revealed the presence of several discrete truncated species that shared a common 5' end and appeared to be intermediates of degradation. S1 mapping showed that the 3' ends of these molecules were distributed throughout the 3' untranslated region, preferentially in A + U-rich regions, broadly correlating with the distribution of AUUUA sites. Nuclear run-on experiments indicated that transcriptional induction accounted for less than 15% of the accumulation of MONAP mRNA. This mRNA was induced in HL60 cells by treatment with several differentiation-inducing agents: 12-O-tetradecanoyl phorbol-13-myristate alone, sodium butyrate, vitamin D3, and dimethyl sulfoxide. It was also induced in quiescent diploid lung fibroblasts stimulated to divide by serum, and it was constitutively overexpressed by some human tumor lines.

scholarly journals Regulation of the mRNA for monocyte-derived neutrophil-activating peptide in differentiating HL60 promyelocytes.

1989 ◽  
Vol 9 (5) ◽  
pp. 1946-1957 ◽  
Author(s):  
J Kowalski ◽  
D T Denhardt
Keyword(s):  
Sodium Butyrate ◽  
Mrna Stability ◽  
Untranslated Region ◽  
Actinomycin D ◽  
Human Tumor ◽  
Lung Fibroblasts ◽  
Macrophage Differentiation ◽  
Cytoplasmic Rna ◽  
Transcriptional Induction ◽  
Full Length Clone

A cDNA library was constructed from HL60 human promyelocyte poly(A)+ RNA harvested 3 h after induction of macrophage differentiation with 12-O-tetradecanoyl phorbol-13-acetate in the presence of cycloheximide. We isolated from this library a 1.6-kilobase full-length clone designated b4 whose corresponding mRNA was greatly increased in abundance in cytoplasmic RNA under these conditions. Dideoxy sequencing revealed that this mRNA encoded MONAP (monocyte-derived neutrophil-activating peptide), a 10-kilodalton monokine with neutrophil-specific chemotactic and enzyme-releasing activities. The 3' untranslated region of this mRNA was found to be 1.2 kilobases long and possessed nine copies of the AUUUA sequence known to be associated with regulation of mRNA stability. Actinomycin D chase experiments yielded evidence that cytoplasmic stabilization was one of the means of regulation of MONAP expression. Analysis of cytoplasmic poly(A)- RNA revealed the presence of several discrete truncated species that shared a common 5' end and appeared to be intermediates of degradation. S1 mapping showed that the 3' ends of these molecules were distributed throughout the 3' untranslated region, preferentially in A + U-rich regions, broadly correlating with the distribution of AUUUA sites. Nuclear run-on experiments indicated that transcriptional induction accounted for less than 15% of the accumulation of MONAP mRNA. This mRNA was induced in HL60 cells by treatment with several differentiation-inducing agents: 12-O-tetradecanoyl phorbol-13-myristate alone, sodium butyrate, vitamin D3, and dimethyl sulfoxide. It was also induced in quiescent diploid lung fibroblasts stimulated to divide by serum, and it was constitutively overexpressed by some human tumor lines.

scholarly journals Identification of a Human VPF/VEGF 3′ Untranslated Region Mediating Hypoxia-induced mRNA Stability

1998 ◽  
Vol 9 (2) ◽  
pp. 469-481 ◽  
Author(s):  
Kevin P. Claffey ◽  
Shu-Ching Shih ◽  
Andrew Mullen ◽  
Suzan Dziennis ◽  
Jennifer L. Cusick ◽  
...  
Keyword(s):  
Mrna Stability ◽  
Untranslated Region ◽  
Stability Region ◽  
Rna Binding ◽  
Malignant Tumors ◽  
Human Tumor ◽  
Luciferase Reporter ◽  
Vegf Expression ◽  
Vegf Mrna ◽  
Vascular Permeability Factor

Hypoxia is a prominent feature of malignant tumors that are characterized by angiogenesis and vascular hyperpermeability. Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) has been shown to be up-regulated in the vicinity of necrotic tumor areas, and hypoxia potently induces VPF/VEGF expression in several tumor cell lines in vitro. Here we report that hypoxia-induced VPF/VEGF expression is mediated by increased transcription and mRNA stability in human M21 melanoma cells. RNA-binding/electrophoretic mobility shift assays identified a single 125-bp AU-rich element in the 3′ untranslated region that formed hypoxia-inducible RNA-protein complexes. Hypoxia-induced expression of chimeric luciferase reporter constructs containing this 125-bp AU-rich hypoxia stability region were significantly higher than constructs containing an adjacent 3′ untranslated region element without RNA-binding activity. Using UV-cross-linking studies, we have identified a series of hypoxia-induced proteins of 90/88 kDa, 72 kDa, 60 kDa, 56 kDa, and 46 kDa that bound to the hypoxia stability region element. The 90/88-kDa and 60-kDa species were specifically competed by excess hypoxia stability region RNA. Thus, increased VPF/VEGF mRNA stability induced by hypoxia is mediated, at least in part, by specific interactions between a defined mRNA stability sequence in the 3′ untranslated region and distinct mRNA-binding proteins in human tumor cells.

AU-rich elements in the mRNA 3′-untranslated region of the rat receptor for advanced glycation end products and their relevance to mRNA stability

2004 ◽  
Vol 319 (1) ◽  
pp. 247-255 ◽  
Author(s):  
José Juan Caballero ◽  
Marı́a Dolores Girón ◽  
Alberto Manuel Vargas ◽  
Natalia Sevillano ◽  
Marı́a Dolores Suárez ◽  
...  
Keyword(s):  
Advanced Glycation End Products ◽  
Mrna Stability ◽  
Untranslated Region ◽  
Advanced Glycation ◽  
Glycation End Products ◽  
End Products

scholarly journals Antiproliferative activity of essential oils from three plants of the Brazilian Cerrado: Campomanesia adamantium (Myrtaceae), Protium ovatum (Burseraceae) and Cardiopetalum calophyllum (Annonaceae)

2020 ◽  
Vol 80 (2) ◽  
pp. 290-294 ◽  
Author(s):  
C. C. F. Alves ◽  
J. D. Oliveira ◽  
E. B. B. Estevam ◽  
M. N. Xavier ◽  
H. D. Nicolella ◽  
...  
Keyword(s):  
Essential Oils ◽  
Cell Line ◽  
Tumor Cells ◽  
Antiproliferative Activity ◽  
Biological Activities ◽  
Chemical Constituents ◽  
Human Tumor ◽  
Lung Fibroblasts ◽  
Breast Adenocarcinoma ◽  
Brazilian Cerrado

Abstract Essential oils, which may be extracted from several parts of plants, have different biological activities. The Brazilian Cerrado has a large variety of plants that yield essential oils, even though many have not been studied yet. Taking into account the biodiversity of this biome, this study aimed at evaluating the antiproliferative activity of essential oils extracted from three species of plants of the Cerrado in Goiás state: Campomanesia adamantium (Cambess.) O. Berg, Protium ovatum (Engl. in Mart.) and Cardiopetalum calophyllum (Schltdl.). Essential oils were extracted from both C. adamantium and C. calophyllum leaves and from P. ovatum leaves and green fruits by hydrodistillation carried out by a Clevenger-type apparatus. The chemical composition of the essential oils was determined by Gas Chromatography coupled to Mass Spectrometry (GC-MS). The following major chemical constituents were identified in the essential oils under investigation: β-myrcene (62.00%), spathulenol (28.78%), germacrene-B (18.27%), β-caryophyllene oxide (16.40%), β-caryophyllene (14.00%), α-pinene (11.30%), viridiflorol (9.99%), limonene (7.30%) and (Z,E)-pharnesol (6.51%). The antiproliferative activity was evaluated in different human tumor cell lines: breast adenocarcinoma (MCF-7), cervical adenocarcinoma (HeLa) and glioblastoma (M059J). A normal human cell line was included (GM07492A, lung fibroblasts). Results showed that essential oils from C. adamantium leaves got the lowest values of IC50 in all strains of tumor cells under evaluation. They were significantly lower than the ones of the normal cell line, an evidence of selectivity. It is worth mentioning that this is the first report of the antiproliferative activity of essential oils from C. adamantium , P. ovatum and C. calophyllum against human tumor cells.

scholarly journals A U-Rich Element in the 5′ Untranslated Region Is Necessary for the Translation of p27 mRNA

2000 ◽  
Vol 20 (16) ◽  
pp. 5947-5959 ◽  
Author(s):  
S. Sean Millard ◽  
Anxo Vidal ◽  
Maurice Markus ◽  
Andrew Koff
Keyword(s):  
Cell Cycle ◽  
Mrna Stability ◽  
Untranslated Region ◽  
Mrna Processing ◽  
Differentiated Cells ◽  
Specific Subset ◽  
Number Of Factors ◽  
Rnp Complex ◽  
Cycle Dependent

ABSTRACT Increased translation of p27 mRNA correlates with withdrawal of cells from the cell cycle. This raised the possibility that antimitogenic signals might mediate their effects on p27 expression by altering complexes that formed on p27 mRNA, regulating its translation. In this report, we identify a U-rich sequence in the 5′ untranslated region (5′UTR) of p27 mRNA that is necessary for efficient translation in proliferating and nonproliferating cells. We show that a number of factors bind to the 5′UTR in vitro in a manner dependent on the U-rich element, and their availability in the cytosol is controlled in a growth- and cell cycle-dependent fashion. One of these factors is HuR, a protein previously implicated in mRNA stability, transport, and translation. Another is hnRNP C1 and C2, proteins implicated in mRNA processing and the translation of a specific subset of mRNAs expressed in differentiated cells. In lovastatin-treated MDA468 cells, the mobility of the associated hnRNP C1 and C2 proteins changed, and this correlated with increased p27 expression. Together, these data suggest that the U-rich dependent RNP complex on the 5′UTR may regulate the translation of p27 mRNA and may be a target of antimitogenic signals.

Progesterone together with estradiol promotes luteinizing hormone β-subunit mRNA stability in rat pituitary cells cultured in vitro

1996 ◽  
Vol 134 (2) ◽  
pp. 236-242 ◽  
Author(s):  
Deokbae Park ◽  
Minseok cheon ◽  
Changmee Kim ◽  
Kyungjin Kim ◽  
Kyungza Ryu
Keyword(s):  
Mrna Stability ◽  
Actinomycin D ◽  
Pituitary Cells ◽  
Mrna Levels ◽  
Ovarian Steroids ◽  
Subunit Mrna ◽  
Rat Pituitary ◽  
Lh Release ◽  
Rat Pituitary Cells

Park D, Cheon M, Kim C, Kim K, Ryu K. Progesterone together with estradiol promotes luteinizing hormoneβ-subunit mRNA stability in rat pituitary cells in vitro. Eur J Endocrinol 1996;134:236–42. ISSN 0804–4643 The present study examined the role of ovarian steroids, estradiol and/or progesterone in the regulation of luteinizing hormone β-subunit (LH-β) mRNA levels and LH release in the rat anterior pituitary cells cultured in vitro. When estradiol (10 nmol/l and/or progesterone (100 nmol/l) were added to the cultures, neither estradiol or progesterone nor both together altered the basal LH-β mRNA levels or LH release. Continuous exposure to gonadotropin-releasing hormone (GnRH, 0.2 nmol/l) for 24 h markedly induced LH-β mRNA accumulation, and in this experimental condition, progesterone alone and progesterone + estradiol further augmented GnRH-induced LH-β mRNA levels and LH release. Then we explored further the possibility that ovarian steroids are involved in modulating LH-β mRNA stability in cultured rat pituitary cells where transcription was inhibited by actinomycin D. Anterior pituitary cells were preincubated with GnRH (0.2 nmol/l) for 16 h and, after removing GnRH from culture medium, the cells were incubated further in the presence of actinomycin D (5 μmol/l) for 24 h. The LH-β mRNA levels gradually declined to about 30% of the control values (zero time point after GnRH removal) in a time-dependent manner. During this period, either progesterone alone or progesterone + estradiol clearly blocked the degradation of LH-β mRNA species. These results indicate that ovarian steroids promote LH-β mRNA stability, thereby contributing to the maintenance of GnRH-stimulated LH-β mRNA levels. Kyungza Ryu, Department of Pharmacology, College of Medicine, Yonsei University, 120-749, Seoul, Korea

scholarly journals Deadenylation-Dependent and -Independent Decay Pathways for α1-Tubulin mRNA in Chlamydomonas reinhardtii

1998 ◽  
Vol 18 (3) ◽  
pp. 1498-1505 ◽  
Author(s):  
Joseph F. Gera ◽  
Ellen J. Baker
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Northern Blot ◽  
Untranslated Region ◽  
Northern Blot Analysis ◽  
Actinomycin D ◽  
Degradation Pathway ◽  
Transformed Cells ◽  
Tubulin Mrna ◽  
Single Transcript ◽  
Β Tubulin

ABSTRACT The α- and β-tubulin mRNAs of Chlamydomonas reinhardtii exhibit different half-lives under different conditions: when expressed constitutively, they degrade with half-lives of about 1 h, whereas when induced by deflagellation, they degrade with half-lives of only 10 to 15 min. To investigate the decay pathway(s) used under these two conditions, an α1-tubulin gene construct which included an insert of 30 guanidylate residues within the 3′ untranslated region was introduced into cells. This transgene was efficiently expressed in stably transformed cells, and the mRNA exhibited constitutive and postinduction half-lives like those of the α1-tubulin mRNA. Northern blot analysis revealed the occurrence of a 3′ RNA fragment derived from the poly(G)-containing α1-tubulin transcripts. The 3′ fragment was shown to accumulate as full-length mRNA disappeared in actinomycin D-treated cells, indicating a precursor-product relationship. Insertion of a second poly(G) tract upstream of the first resulted in accumulation of only a longer 3′ fragment, suggesting that the decay intermediate is generated by 5′-to-3′ exonucleolytic digestion. A translational requirement for generation of the 3′ fragment was demonstrated by experiments in which cells were deflagellated in the presence of cycloheximide. Analysis of fragment poly(A) length revealed that the fragments were, at most, oligoadenylated in nondeflagellated cells but had a long poly(A) tail in deflagellated cells. These findings suggest that the oligoadenylated fragment is a decay intermediate in a deadenylation-dependent, constitutive degradation pathway and that the requirement for deadenylation is bypassed in deflagellated cells. This represents the first example in which a single transcript has been shown to be targeted to different decay pathways under different cellular conditions.

Hypoxia downregulates tropoelastin gene expression in rat lung fibroblasts by pretranslational mechanisms

1999 ◽  
Vol 277 (3) ◽  
pp. L566-L572 ◽  
Author(s):  
John L. Berk ◽  
Nima Massoomi ◽  
Christine Hatch ◽  
Ronald H. Goldstein
Keyword(s):  
Gene Expression ◽  
Gene Transcription ◽  
Actinomycin D ◽  
Lung Fibroblasts ◽  
Mrna Levels ◽  
Rat Lung ◽  
Cell Toxicity ◽  
Chromium Release ◽  
Injured Lung ◽  
Isolated Rat Lung

Elastolytic lung injury disrupts cell barriers, flooding alveoli and producing regional hypoxia. Abnormal O2 tensions may alter repair of damaged elastin fibers. To determine the effect of hypoxia on extravascular elastin formation, we isolated rat lung fibroblasts and cultured them under a variety of O2 conditions. Hypoxia downregulated tropoelastin mRNA in a dose- and time-related fashion while upregulating glyceraldehyde-3-phosphate dehydrogenase mRNA levels. The changes in tropoelastin gene expression were not due to cell toxicity as measured by chromium release and cell proliferation studies. Neither cycloheximide nor actinomycin D abrogated this effect. Hypoxia induced early decreases in tropoelastin mRNA stability; minor suppression of gene transcription occurred later. When returned to 21% O2, tropoelastin mRNA recovered to control levels in part by upregulating tropoelastin gene transcription. Taken together, these data indicate that hypoxia regulates tropoelastin gene expression and may alter repair of acutely injured lung.

Robust induction of PGHS-2 by IL-1 in orbital fibroblasts results from low levels of IL-1 receptor antagonist expression

2003 ◽  
Vol 284 (6) ◽  
pp. C1429-C1437 ◽  
Author(s):  
H. James Cao ◽  
Rui Han ◽  
Terry J. Smith
Keyword(s):  
Receptor Antagonist ◽  
Proinflammatory Cytokines ◽  
Mrna Stability ◽  
Untranslated Region ◽  
Promoter Activity ◽  
Gene Promoter ◽  
Transcript Stability ◽  
Low Levels ◽  
Endoperoxide H Synthase ◽  
Orbital Fibroblasts

Human orbital fibroblasts are more susceptible to some actions of proinflammatory cytokines than are fibroblasts from other anatomic regions. These cells produce high levels of PGE2when activated by cytokines. Here we report that they express high levels of prostaglandin-endoperoxide H synthase (PGHS)-2, the inflammatory cyclooxygenase, when treated with IL-1β. This induction results from enhanced PGHS-2 mRNA stability and small increases in gene promoter activity. The enhanced transcript stability is a result of actions of the cytokine on the 3′-untranslated region. Orbital fibroblasts, unlike those from skin, fail to express high levels of IL-1 receptor antagonist (IL-1ra) when treated with IL-1β, leading to loss of modulation of IL-1 action. This can be overcome by transiently transfecting cells with IL-1ra. Thus a decreased level of IL-1ra expression in orbital fibroblasts may underlie the exaggerated responses to IL-1 observed in those cells and, therefore, the susceptibility of the orbit to inflammation.

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