共查询到20条相似文献,搜索用时 15 毫秒
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Relationship of c-myc gene copy number and gene expression: cellular effects of elevated c-myc protein 总被引:1,自引:0,他引:1
M G Pallavicini C Rosette M Reitsma P S Deteresa J W Gray 《Journal of cellular physiology》1990,143(2):372-380
The relationship between gene copy number and expression and cellular consequences of elevated levels of c-myc protein has been investigated using recombinant Chinese hamster ovary cell lines transfected with DNA coding for the murine c-myc gene. HC-8 and LC-5 recombinant cells carry approximately 800 and 50 copies of c-myc sequences, respectively, under control of an inducible heat shock promoter. Multivariate flow cytometric analysis and clonogenic assays were used to measure the relationship among c-myc expression, rate of DNA synthesis, and cell survival. Following heat exposure, maximally induced HC-8 cells produced approximately tenfold more c-myc protein than heated LC-5 cells, suggesting a close relationship between gene copy number and level of expression. However, considerable heterogeneity in the level and time of c-myc expression was observed following heat induction, even though the amounts of genomic c-myc were relatively constant. Heterogeneity in gene expression was not attributable to variation in heat induction methodologies and/or cell cycle phase distributions. The presence of high levels of recombinant c-myc protein was associated with a decreased rate of bromodeoxyuridine incorporation into DNA. High levels of c-myc protein in HC-8 cells were inversely correlated with cell survival postheating, suggesting that high levels of c-myc protein are incompatible with cell survival. 相似文献
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Genome-scale compositional analyses of non-coding sequences from 410 microbes of varying GC-content, lineage, environment/life-style, reveal presence of a distinct trend in GC-usage in spacers between intra-operonic and extra-operonic gene-pairs. For most of the microbes, average GC-content of the intra-operonic spacers are consistently higher than those between extra-operonic unidirectional gene-pairs. Also, unidirectional gene-pairs exhibiting higher cross-species conservation, irrespective of their operonic context, house relatively GC-rich spacers. A few prokaryotes, most of which represent known cases of genome degradation, stand out as exceptions defying this trend. GC-enrichment of intra-operonic spacers therefore appears to be an evolutionary strategy facilitating preservation of operonic gene-order. 相似文献
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Binding of tryptophan to plasma proteins in several species 总被引:2,自引:0,他引:2
R W Fuller B W Roush 《Comparative biochemistry and physiology. B, Comparative biochemistry》1973,46(2):273-276
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W. Keith Miskimins 《Journal of cellular biochemistry》1992,49(4):349-356
Nuclear factors from HeLa cells were isolated by elution of DNA-cellulose bound proteins with a double stranded synthetic oligonucleotide corresponding to the region from ?34 to ?79 of the human transferrin receptor (TR) gene promoter. The eluted proteins were further purified and separated from the oligonucleotide by ion exchange chromatography. Proteins within the resulting fraction bound with specificity to the TR promoter. Retardation gel analysis and competition with specific double-stranded oligonucleotides show that multiple factors present in this fraction compete for binding within the same region of the TR promoter. Footprinting experiments demonstrate that these factors contact a GC-rich element that is within the region that is required for enhanced expression of the gene in proliferating cells. One of the factors protects an extended DNA sequence but, still contacts the GC-rich element. 相似文献
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O K Gasymov A R Abduragimov T N Yusifov B J Glasgow 《Biochimica et biophysica acta》1999,1433(1-2):307-320
The principal lipid binding protein in tears, tear lipocalin (TL), binds acid and the fluorescent fatty acid analogs, DAUDA and 16-AP at one site TL compete for this binding site. A fluorescent competitive binding assay revealed that apo-TL has a high affinity for phospholipids and stearic acid (Ki) of 1.2 microM and 1.3 microM, respectively, and much less affinity for cholesterol (Ki) of 15.9 of the hydrocarbon chain. TL binds most strongly the least soluble lipids permitting these lipids to exceed their maximum solubility in aqueous solution. These data implicate TL in solubilizing and transporting lipids in the tear film. Phenylalanine, tyrosine and cysteine+ were substituted for TRP 17, the only invariant residue throughout the lipocalin superfamily. Cysteine substitution resulted in some loss os secondary structure, relaxation of aromatic side chain rigidity, decreased binding affinity for DAUDA and destabilization of structure. Mutants of TL, W17Y, and W17F showed a higher binding affinity for DAUDA than wild-type TL. Comparison of the results of the tryptophan 17 substitution in lipocalin with those of tryptophan 19 substitution in beta-lactoglobulin revealed important differences in binding characteristics that reflect the functional heterogeneity within the lipocalin family. 相似文献
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The DNA binding and cleavage properties of quercetin nickel (II) complex have been studied, but little attention has been
devoted to the relationship between antitumor activity of this complex and DNA-binding properties. In the present study, we
report that quercetin nickel (II) complex showed significant cytotoxicity against three tumor cell lines (HepG2, SMMC7721
and A549). Hoechst33258 and AO/EB staining showed HepG2 cells underwent the typical morphologic changes of apoptosis characterized
by nuclear shrinkage, chromatin condensation, or fragmentation after exposure to quercetin nickel (II) complex. We also demonstrate
that the levels of survivin and bcl-2 protein expression in HepG2 cells decreased concurrently, and the levels of p53 protein
increased significantly after treatment with quercetin nickel (II) complex by immunocytochemistry analysis. The relative activity
of caspase-3 and caspase-9 increased significantly after treatment with the complex. Furthermore, fluorescence measurements
and molecular modeling were performed to learn that the complex could be preferentially bound to DNA in GC region. These results
imply that quercetin nickel (II) complex may intercalate into the GC-rich core promoter region of survivin, down-regulating
survivin gene expression and promoting tumor cells apoptosis. So our results suggest that antitumor activity of quercetin
nickel (II) complex might be related to its intercalation into DNA and DNA-binding selectivity, and that the complex may be
a promising agent for cancer therapy. 相似文献
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Metabolism of c-myc gene products: c-myc mRNA and protein expression in the cell cycle. 总被引:24,自引:6,他引:18 下载免费PDF全文
P H Rabbitts J V Watson A Lamond A Forster M A Stinson G Evan W Fischer E Atherton R Sheppard T H Rabbitts 《The EMBO journal》1985,4(8):2009-2015
The presence and synthesis of c-myc protein and mRNA in the cell cycle has been studied. We find that c-myc mRNA is present, at equivalent levels, at all times in the cell cycle with the possible exception of mitosis. Furthermore, we demonstrate that this mRNA is transcribed in both G1 and G2 phases. An analysis of the c-myc protein in vivo shows that de novo synthesis occurs in G1 and G2 and the protein turns over with a half-life of approximately 20-30 min in both phases. Furthermore, the level of c-myc protein rapidly increases in cell populations when they re-initiate the cell cycle, thereafter decreasing as the culture reaches quiescence. The results therefore suggest that expression of c-myc can be rapidly modulated and that it is activated during the G0 to G1 transition, but is expressed thereafter in the cell cycle. 相似文献
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Chromatin structure and protein binding in the putative regulatory region of the c-myc gene in Burkitt lymphoma 总被引:41,自引:0,他引:41
A chromosomal myc gene displays one of three patterns of activity depending upon the arrangement of the gene and its allelic partner. In nonmalignant B cells both myc alleles are normally expressed. In Burkitt lymphoma cells carrying both a translocated and a nontranslocated myc allele, the translocated allele is inappropriately expressed, while the nontranslocated allele is virtually inactive. Here we examine the chromatin structure of these genes using DNAase I hypersensitivity in nonmalignant lymphoblastoid cells and in the Burkitt lymphoma, BL31 . Three hypersensitivity patterns emerge that correlate with the state of the gene and reveal sites associated with putative regulatory structures. One region is associated with the two myc promoters, one with a specific nuclear protein binding site, and one--which is markedly enhanced in the inactive germline gene in the Burkitt cell--with a putative negative control region. The perturbation of the normal pattern in this particular Burkitt cell may be due to the action of an immunoglobulin enhancer. 相似文献
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Comparative studies on the regulation of tryptophan synthesis 总被引:11,自引:0,他引:11
I P Crawford 《CRC critical reviews in biochemistry》1980,8(2):175-189
In vitro DNA recombination techniques have revolutionized the study of genetic control of biosynthetic pathways. Using examples drawn from the pathway of tryptophan synthesis, approaches to the deciphering of regulatory signals and response mechanisms through transposition of DNA segments and DNA sequence analysis will be presented. After reviewing the known chromosomal arrangements and regulatory patterns of trp genes in the bacterial groups studied so far, and describing the results of transferring all or part of the pathway's genes from one organism to a distantly related one, the use of this technique to analyze new organisms will be described. Along with some advantages over the conventional methods there are some pitfalls. Finally, since it is likely that events analogous to recombinant DNA experiments take place readily in nature, their consequences in studies of bacterial evolution will be conjectured. 相似文献
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Apolipoprotein A-I (apoAI) contains several amphipathic α-helices. To carry out its function, it exchanges between lipid-free and different lipidated states as bound to membranes or to lipoprotein complexes of different morphology, size, and composition. When bound to membranes or to spherical lipoprotein surfaces, it is thought that most α-helices arrange with their long axis parallel to the membrane surface. However, we previously found that a central region spanning residues 87-112 is exclusively labeled by photoactivable reagents deeply located into the membrane (Co?rsico et al. (2001) J. Biol. Chem. 276, 16978-16985). A pair of amphipathic α-helical repeats with a particular charge distribution is predicted in this region. In order to study their insertion topology, three single tryptophan mutants, each one containing the tryptophan residue at a selected position in the hydrophobic face of the central Y-helices (W@93, W@104, and W@108), were used. From the accessibility to quenchers located at different membrane depths, distances from the bilayer center of 13.4, 10.5, and 15.7 ? were estimated for positions 93, 104, and 108, respectively. Reported data also indicate that distances between homologous positions (in particular for W@93 and W@104) are very short in dimers in aqueous solution, but they are larger in membrane-bound dimers. Data indicate that an intermolecular central Y-helix bundle would penetrate the membrane perpendicularly to the membrane surface. Intermolecular helix-helix interactions would occur through the hydrophilic helix faces in the membrane-bound bundle but through the hydrophobic faces in the case of dimers in solution. 相似文献
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S. Arhondakis M. Milanesi T. Castrignanò S. Gioiosa A. Valentini G. Chillemi 《Animal genetics》2020,51(3):358-368
Vertebrate genomes are mosaics of megabase-size DNA segments with a fairly homogeneous base composition, called isochores. They are divided into five families characterized by different guanine-cytosine (GC) levels and linked to several functional and structural properties. The increased availability of fully sequenced genomes allows the investigation of isochores in several species, assessing their level of conservation across vertebrate genomes. In this work, we characterized the isochores in Bos taurus using the ARS-UCD1.2 genome version. The comparison of our results with the well-studied human isochores and those of other mammals revealed a large conservation in isochore families, in number, average GC levels and gene density. Exceptions to the established increase in gene density with the increase in isochores (GC%) were observed for the following gene biotypes: tRNA, small nuclear RNA, small nucleolar RNA and pseudogenes that have their maximum number in H2 and H1 isochores. Subsequently, we assessed the ontology of all gene biotypes looking for functional classes that are statistically over- or under-represented in each isochore. Receptor activity and sensory perception pathways were significantly over-represented in L1 and L2 (GC-poor) isochores. This was also validated for the horse genome. Our analysis of housekeeping genes confirmed a preferential localization in GC-rich isochores, as reported in other species. Finally, we assessed the SNP distribution of a bovine high-density SNP chip across the isochores, finding a higher density in the GC-rich families, reflecting a potential bias in the chip, widely used for genetic selection and biodiversity studies. 相似文献
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The optimal parameters in the use of nuclease S1 in DNA reassociation kinetics in the presence of formamide have been determined. The conditions are especially suitable for the study of DNA rich in mole percent GC. A 10-fold dilution of the reassociation samples leading to a decrease in both NaCl and formamide concentrations, consequently resulting in a lowering of Tm by only 1.5 degree C, and the S1 digestion at temperatures identical to the reassociation assay in order to retain the stability of the duplex, are two important aspects of this system. Under these conditions, the kinetics of reassociation followed the theoretically predicted pattern, while the earlier reported methods have shown lower values. 相似文献
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