Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have investigated the role of the insulin-like growth factor one (IGF1) receptor and its relationship to the proto-oncogene c-myb in the growth of two types of hemopoietic cells: mitogen-stimulated human peripheral blood mononuclear cells and a human promyelocytic cell line (HL-60). Using the antisense strategy and the reverse transcriptase polymerase chain reaction (RT-PCR), we show that expression of the IGF1 receptor is required for the entry into S phase of both stimulated lymphocytes and HL-60 cells. The inhibition of DNA synthesis by antisense oligomers to the IGF1 receptor RNA is accompanied by an inhibition of the expression of the mRNA for a DNA synthesis gene, proliferating cell nuclear antigen (PCNA), the co-factor of DNA polymerase delta. Inhibition of c-myb expression results in a decrease in IGF1 receptor mRNA levels; on the other hand, inhibition of IGF1 receptor expression has no effect on c-myb mRNA levels. A tentative temporal relationship between these three genes (c-myb, IGF1 receptor, PCNA) is proposed.
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PMID:The insulin-like growth factor 1 receptor is required for the proliferation of hemopoietic cells. 135 94

Breast tumor cell lines have been shown to secrete at least five distinct insulin-like growth factor (IGF) binding proteins (IGFBP), the secretion being related to the estrogen receptor (ER) content. In this study we investigated IGFBP mRNA expression and IGFBP content in relation to ER content in human breast tumors. Tissue specimens from 47 breast cancers were studied. In five cases the adjacent histologically normal tissue was also analyzed. IGFBP content in tissue homogenates was studied by Western ligand blot analysis, using [125I] IGF-I as a label, and IGFBP mRNA expression by reverse transcriptase polymerase chain reaction. The results show that human breast tumors express mRNAs encoding IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4 and IGFBP-5. The pattern of IGFBPs in different tumors varies. No correlation exists between ER content and IGFBPs with molecular weights 24,000 Mr, 28,000 Mr, 34,000 Mr or 43,000 Mr, whereas the 49,000 Mr IGFBP was more abundant in ER negative tumors (P less than 0.05). The IGFBP content was significantly (P less than 0.05) higher in five tumors than in their adjacent normal tissues suggesting that increased content of IGFBPs is a feature typically associated with the malignant transformation of breast tissue.
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PMID:Insulin-like growth factor binding proteins in human breast cancer tissue. 138 38

The insulin-like growth factor (IGF)-II/mannose-6-phosphate (M6P) receptor, which targets acid hydrolases to lysosomes, is a multifunctional protein with separate binding sites for IGF-II and M6P. The purpose of this study was to determine if alveolar macrophages (AM) and their precursor cells, blood monocytes, expressed this receptor. AM expressed IGF-II/M6P receptors as detected by [125]IGF-II surface binding that was not reduced by recombinant IGF-I or IGF-I receptor monoclonal antibody (alpha IR3). Surface binding was also detected on blood monocytes and could be upregulated approximately 4-fold by incubation with lipopolysaccharide. There were no differences in surface binding by AM lavaged from individuals with asbestos exposure or from normal volunteers. Using the polymerase chain reaction and reverse transcriptase to reverse-transcribe mRNA from mononuclear phagocytes, specific IGF-II/M6P receptor cDNA was amplified and detected by agarose gel electrophoresis from both AM and blood monocytes. The IGF-II/M6P receptor has an intracellular transport role in many cells cycling from the cell surface to the cytoplasm, or binding to phosphorylated acid hydrolases in the Golgi and transporting them to an acidic prelysosomal site where they dissociate and fuse to the lysosomes and IGF-II/M6P recycles to the trans-Golgi. These functions may be particularly important in asbestosis and other interstitial lung diseases where AM are activated, intracellular lysosomes are a prominent morphologic feature, and acid hydrolases are found in recovered lavage fluid.
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PMID:Human mononuclear phagocytes express the insulin-like growth factor-II/mannose-6-phosphate receptor. 164 80

The insulin-like growth factor (IGF) system is thought to function as a mediator of steroid hormone actions in the endometrium. IGFs (IGF-I and IGF-II) are also potent mitogens in endometrial cancer. The biological actions of IGFs are modulated by specific binding proteins (IGFBP)--6 cloned and sequenced so far--which may either inhibit or enhance the effects of IGF at the cellular level. In the endometrium, IGFBP-1 gene expression is stimulated by progesterone and inhibited by insulin, while IGFBP-1 inhibits the mitogenic action of IGF-I. In this study, we used a quantitative reverse transcriptase polymerase chain reaction (RT-PCR) to investigate IGFBP-1, IGFBP-2, IGFBP-4, IGFBP-5 and IGFBP-6 gene expression in endometrial cancer tissues. Endometrial cancer tissue samples were collected from 20 women (aged 54-79 yrs) with stage I to II well-differentiated endometrial adenocarcinoma. Samples of normal endometrium (n = 14) obtained from women undergoing tubal ligation in various phases of the menstrual cycle, and normal early-pregnancy endometrium (decidua) were studied for comparison. In endometrial cancer tissues, the IGFBP-1 mRNA was undetectable or minimally expressed when studied by RT-PCR. The mean (+ SD) levels of IGFBP-2 and IGFBP-4 and IGFBP-5 mRNAs in endometrial cancer tissues did not differ from those in normal endometrium, in which no cyclic variation was observed, suggesting that the genes encoding IGFBP-2, IGFBP-4 and IGFBP-5 are not hormonally regulated in the endometrium. The IGFBP-6 mRNA expression showed a significant cyclic variation in normal endometrium, with low levels in late-proliferative and early- to mid-secretory phases and high expression in late-secretory and early-proliferative phases. In endometrial cancer tissues, the mean IGFBP-6 mRNA level was similar to that in cycling endometrium during the peri-ovulatory period. In summary, a continuous stimulation of the endometrial epithelial cells by IGFs with suppressed IGFBP-1 expression may lead to an imbalance in the IGF system of the endometrium and trigger an uncontrolled cell proliferation, ultimately resulting in malignant transformation.
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PMID:Suppressed expression of insulin-like growth factor binding protein-1 mRNA in the endometrium: a molecular mechanism associating endometrial cancer with its risk factors. 752 16

Senescent human diploid fibroblasts express several growth-regulated genes but fail to express others. In this paper we show, by a very sensitive technique (reverse transcriptase-polymerase chain reaction), that senescent cells fail to express insulin-like growth factor-1 (IGF-1) mRNA, which is expressed in moderate amounts by young cells. Human fibroblasts immortalized by transfection with a temperature-sensitive SV40 T antigen gene regain the ability to express IGF-1 mRNA, but only at the permissive temperature of 34 degrees C. Under these conditions, the immortalized human fibroblasts grow even in 1% serum. At the restrictive temperature of 39 degrees C, the temperature-sensitive T antigen is nonfunctional, IGF-1 RNA is not detectable, and the cells fail to grow even in 10% serum. The failure to express IGF-1 mRNA in postsenescent cells can be ascribed, at least in part, to a transcriptional mechanism. Despite the correlation among immortalization by SV40 T antigen, expression of IGF-1, and growth, it seems unlikely that the failure to express IGF-1 is the sole cause of cellular senescence; other requirements must be postulated.
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PMID:Failure of senescent human fibroblasts to express the insulin-like growth factor-1 gene. 768 32

SPC2 and SPC3 are two members of a family of subtilisin-related proteases which play essential roles in the processing of prohormones into their mature forms in the pancreatic B cell and many other neuroendocrine cells. To investigate the phylogenetic origins and evolutionary functions of SPC2 and SPC3 we have identified and cloned cDNAs encoding these enzymes from amphioxus (Branchiostoma californiensis), a primitive chordate. The amino acid sequence of preproSPC2 contains 689 aa and is 71% identical to human SPC2. In contrast, amphioxus prproSPC3 consists of 774 aa and exhibits 55% identity to human SPC3. These results suggest that the primary structure of SPC2 has been more highly conserved during evolution than that of SPC3. To further investigate the function(s) of SPC2 and SPC3 in amphioxus, we have determined the regional expression of these genes by using a reverse transcriptase-linked polymerase chain reaction (RT-PCR) assay. Whole amphioxus was dissected longitudinally into four equal-length segments and RNA was extracted. Using RT-PCR to simultaneously amplify SPC2 and SPC3 DNA fragments, we found that the cranial region (section 1) expressed equal amounts of SPC2 and SPC3 mRNAs, whereas in the caudal region (section 4) the SPC2-to-SPC3 ratio was 5:1. In the mid-body sections 2 and 3 the SPC2-to-SPC3 ratio was 1:5. By RT-PCR we also determined that amphioxus ILP, a homologue of mammalian insulin/insulin-like growth factor, was expressed predominately in section 3. These results suggest that the relative levels of SPC2 and SPC3 mRNAs are specifically regulated in various amphioxus tissues. Furthermore, the ubiquitous expression of these mRNAs in the organism indicates that they are involved in the processing of other precursor proteins in addition to proILP.
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PMID:Proprotein convertases in amphioxus: predicted structure and expression of proteases SPC2 and SPC3. 772 4

We have recently demonstrated that rat cytomegalovirus (RCMV) infection induces an early inflammatory response in the adventitia (perivasculitis) and in the subendothelial space (endothelialitis) as well as doubles smooth muscle cell (SMC) proliferation and intimal thickening of rat aortic allografts performed from the DA (AG-B4, RT1a) to the WF (AG-B2, RT1v) strain. In this study, the impact of RCMV infection on the structure of inflammation in the allograft adventitia and on the expression of SMC growth factors in the allograft vascular wall was investigated. The recipient rats were inoculated with 10(5) plaque-forming U of RCMV Maastricht strain or left noninfected and used as controls. The allografts were removed at 7 days and 1 and 3 months after transplantation and processed for morphometry and immunohistochemistry. RNA was isolated for reverse transcriptase polymerase chain reaction (RT-PCR). RCMV infection was associated with significantly upregulated presence (P < .05) of T helper (W3/25), T cytotoxic (OX8), and natural killer (3.2.3) cells in the allograft adventitia 7 days after transplantation but not thereafter. More monocyte/macrophages (OX42) were also present in RCMV-infected allografts, but the difference was not significant. Concomitantly, RCMV infection significantly enhanced (P < .05) the expression of major histocompatibility complex class II (OX6) and almost doubled (P = NS) the expression of interleukin-2R (CD25), intercellular adhesion molecule-1 (CD54;1A29), and lymphocyte function-associated antigen-1 alpha-chain (CD11a; WT.1) in the adventitial inflammatory infiltrate. RCMV infection was linked with an early, prominent expression of both PDGF-BB mRNA at 7 days (P < .05) and at 1 month (P < .025) and of transforming growth factor-beta 1 mRNA at 7 days (P < .025) and at 1 month (P < .025) after transplantation. A less-prominent mRNA upregulation of acidic fibroblast growth factor (P < .05) was associated with RCMV infection at 7 days and at 1 month, as well as of epidermal growth factor at 1 month after transplantation, when compared with noninfected allografts, although the mRNA expression in both groups was below the levels of nontransplanted DA aortas. RCMV infection almost doubled basic fibroblast growth factor mRNA expression (P = NS) in the allograft vascular wall at 7 days and at 1 month. RCMV infection had no additional impact on insulin-like growth factor-1 mRNA expression when compared with noninfected allografts.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Cytomegalovirus infection enhances mRNA expression of platelet-derived growth factor-BB and transforming growth factor-beta 1 in rat aortic allografts. Possible mechanism for cytomegalovirus-enhanced graft arteriosclerosis. 798 Nov 94

The ovine insulin-like growth factor-II (oIGF-II) gene is comprised of 9 exons that span approximately 25 kb. Approximately 750 nucleotides upstream of oIGF-II exon 1 are the three exons of the ovine insulin gene that are transcribed in the same direction as oIGF-II. The genomic organization and expression of the oIGF-II gene is similar to that of the human IGF-II gene. Four putative promoters direct the transcription of six 5' noncoding exons (1, 3, 4, 5, 6, and 7), which are alternatively spliced to the shared exons 8, 9, and 10. An ovine exon comparable to human exon 2 has not been identified. Multiple transcription initiation sites were identified for exons 1 and 6 by primer extension analysis. Using a reverse transcriptase polymerase chain reaction (RT-PCR) assay, exon 1 and 3 transcripts were shown to be expressed in adult but not fetal liver. In addition, a novel transcript, which contained exon 1 spliced directly to exon 8, was detected in adult liver. Exon 4 transcripts were not detected in either fetal or adult liver, whereas exon 6 and 7 transcripts were detected in both fetal and adult liver. Exon 5 transcripts were also expressed in both fetal and adult liver, which is in contrast to the tumor cell-specific expression of human exon 5. Like the human and rodent genes, the regulation of expression of the oIGF-II gene is under complex control.
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PMID:Characterization of the linked ovine insulin and insulin-like growth factor-II genes. 801 Nov 64

To address the question whether fish brain can produce insulin, pink salmon (Oncorhynchus gorbusha) brains were extracted and processed according to the procedure developed for purification of pancreatic insulin (Rusakov and Bondareva, 1979). Biological and immunological activity of the resulting material was evaluated respectively by a cartilage sulfation assay and by radioimmunoassay homologous for salmon insulin. Preparations from salmon brain stimulated the [35S]sulfate uptake into salmon branchial cartilage with a potency comparable to pure mammalian or salmon insulins but lower than that of mammalian insulin-like growth factor (IGF-I). In contrast, only trace amounts of radioimmunoreactive insulin could be detected by homologous radioimmunoassay. To determine whether insulin mRNA was present in salmon brain, primers specific for salmon proinsulin and salmon prepro-IGF-I were designed to amplify corresponding cDNA regions by reverse transcriptase-PCR. Insulin mRNA was found only in the endocrine pancreas (Brockmann body) while IGF-I mRNA was detected in the brain, liver, and the Brockmann body. Our results suggest that in fish pancreatic-type insulin is most likely produced only in the endocrine pancreas and then transported to the brain through blood/cerebrospinal fluid system. However, it does not exclude a possibility that some yet unknown insulin-like substances may be expressed in the neural system of ectotherm vertebrates.
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PMID:Does salmon brain produce insulin? 840 93

Messenger RNAs (mRNA) of several growth factor receptors and relate genes were examined with reverse transcriptase polymerase chain reaction (RT-PCR) in normal and noise-damaged chicken basilar papillae (BP). Analysis of the amplification products indicated the presence of mRNAs for epidermal growth factor receptor (EGFR), fibroblast factor receptor (FGFR), insulin-like growth factor receptor (IGFR), insulin receptor (IR), retinoic acid receptor beta (RAR beta), retinoic acid receptor gamma (RXR gamma), and basic fibroblast growth factor (BFGF) in both normal and noise-damaged BP. The RT-PCR products generated were characterized by size and sequencing analysis to confirm the identities of the target molecules. The subcellular localization of the mature protein analogs for EGFR, FGFR, IGFR, RAR beta, and BFGF were identified using fluorescence immunocytochemistry and confocal laser scanning microscopy. These experiments indicated that EGFR is present in the stereociliary bundles in the hair cells, IGFR is not present in the cells of the BP, BFGF localizes in the nuclei of supporting cells in the BP, but not hair cells or hyaline cells, and that RAR beta localizes in the perinuclear regions of hair cells. The subcellular distributions of these proteins were consistent in both noise-damaged and control BP. FGFR, in contrast, changed its distribution in the tissue after noise damage. In normal BP, FGFR is concentrated in the stereocilia of hair cells. However, in damaged regions of noise-exposed chick cochleae, FGFR is heavily expressed in the expanded apical regions of the supporting cells. These findings suggest that BFGF and retinoic acid may potentially play a role in the mechanisms which regulate the regeneration of chicken cochlear hair cells.
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PMID:Potential role of bFGF and retinoic acid in the regeneration of chicken cochlear hair cells. 878 6


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