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Functional Analysis Of The Transcriptional Enhancer Of A Chicken U1 Small Nuclear Rna Gene
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Book Synopsis Functional Analysis of the Transcriptional Enhancer of a Chicken U1 Small Nuclear RNA Gene by : Kenneth Allan Roebuck
Download or read book Functional Analysis of the Transcriptional Enhancer of a Chicken U1 Small Nuclear RNA Gene written by Kenneth Allan Roebuck and published by . This book was released on 1989 with total page 296 pages. Available in PDF, EPUB and Kindle. Book excerpt: The DNA sequence requirements of chicken U1 RNA gene expression have been examined in a Xenopus laevis oocyte transcription system. A transcriptional enhancer, which is required for efficient U1 RNA gene expression, extends over approximately 50 base pairs of conserved DNA sequence located 180 to 230 base pairs upstream of the U1 transcription initiation site. It is composed of multiple functional motifs, including a GC-box, an octamer motif, and a novel SPH motif, that contribute synergistically to U1 enhancer activity. Under non-competitive conditions in oocytes, the SPH motif is capable of stimulating U1 RNA transcription in the absence of the other functional motifs, whereas the GC-box and octamer motifs lack this ability. However, in order to form a transcription complex that is stable to challenge by a second competing snRNA transcription unit, both the octamer and SPH motifs are required. The GC-box, although required for full enhancer activity, is not essential for stable complex formation in oocytes. Site-directed mutagenesis of the U1 enhancer was used to study the DNA sequence requirements of the SPH motif. Functional activity of the SPH motif is spread throughout a 24 base pair region immediately 3$\sp\prime$ of the octamer but is particularly dependent upon sequences near an SphI restriction endonuclease site located at the center of the SPH motif. The ability of an SPH-specific factor (SBF) to recognize and bind to mutant enhancer sequences in vitro correlates with the functional activity of the corresponding DNA sequences in vivo. Although the octamer binding factor and SBF can bind independently to their recognition motifs, the formation of a stable transcription complex appears to be dependent upon the cooperative action of these two factors bound to adjacent sites in the enhancer region. Multiple sequence motifs, which are recognized by distinct factors and can function in a variety of cis-linked configurations, appear to be a general structural feature of vertebrate small nuclear RNA gene enhancers and may provide a mechanism to ensure that these housekeeping-type genes are adequately expressed in a wide variety of cell-types.
Book Synopsis Analysis of SBF, an SnRNA Enhancer Binding Protein, and Cloning of the Chicken Rreb-1 Gene by : Jon Hatsuo Miyake
Download or read book Analysis of SBF, an SnRNA Enhancer Binding Protein, and Cloning of the Chicken Rreb-1 Gene written by Jon Hatsuo Miyake and published by . This book was released on 1997 with total page 332 pages. Available in PDF, EPUB and Kindle. Book excerpt: Vertebrate snRNA genes contain two well conserved regions in their 5' flanking DNA sequence. The distal sequence element serves as an enhancer to stimulate gene expression and the proximal sequence element functions to specify the transcription start site. The two U4 genes in the chicken genome code for distinct sequence variants of U4 snRNA that are differentially expressed during development. Whereas U4B RNA is constitutively expressed, U4X RNA is specifically down-regulated relative to U4B in a tissue-specific manner during development. The distal sequence elements for the U1, U4B and U4X snRNA genes contain an octamer binding site juxtaposed to an SPH motif which has been shown to bind the SBF factor. In Chapter 1, I describe the analysis of protein-DNA interactions at the promoter and enhancer regions of chicken U4B and U4X genes. Both genes are expressed at similar levels in the early embryo but U4X expression is specifically down-regulated relative to U4B as development proceeds. I demonstrate that the SPH binding factor, SBF, binds to the U4X SPH motif 20-30 fold less efficiently than to the SPH motif of the U4B gene. I also demonstrate that the Oct-1 factor has similar affinities for the octamer sequence of both genes. Binding assays using nuclear extracts from various developmental stages and tissues revealed that Oct-1 and SBF protein levels do not change throughout development or in different tissues. In contrast, a proximal sequence factor termed PPBF that only binds to the promoter of the U4X gene is up-regulated during development and varies between tissues. Overall, these observations suggest potential roles for SBF and PPBF in the preferential down regulation of U4X gene expression. In chapter 2 I further analyze the properties of the SBF factor. First I address if the Oct-1 and SBF factors can cooperatively bind to the U4B enhancer region. Earlier studies in our lab indicated that SBF and Oct-1 function synergistically in transcription of the U1 and U4B enhancers. However, electrophoretic mobility shift studies indicate the synergistic effect between these factors is not caused by cooperative DNA binding in vitro. I also report that similar SBF-like activities are found in Xenopus oocyte and HeLa cell nuclear extracts. Regarding the chicken and HeLa SBF factor, UV cross-linking studies demonstrate that both are approximately 85 kDa. In chapter 3 I report attempts to clone the SBF gene from a chicken embryo cDNA library and the fortuitous cloning and molecular characterization of the gene encoding the chicken homologue to human RREB-1. Preliminary studies indicated that a candidate clone for SBF had been obtained but isolation of further cDNA clones and antibodies directed against the cloned gene determined that this gene did not encode SBF. Near the completion of this project, other investigators identified a gene in a human medullary thyroid carcinoma cell line termed rreb-1 that is 53% identical and 69% similar to the deduced amino acid sequence of the gene I cloned. Amino acid alignment and the arrangement and sequence of the zinc fingers suggest that the gene I cloned encodes the chicken homologue to the human RREB-1 protein, a Ras-responsive element binding protein. The deduced amino acid sequence for chicken RREB-1 is greatly extended at its N-terminus compared to human RREB-1 and explanations for this are discussed.
Book Synopsis Progress in Molecular and Subcellular Biology by : Philippe Jeanteur
Download or read book Progress in Molecular and Subcellular Biology written by Philippe Jeanteur and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 143 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biological functions are almost exclusively attributed to macromolecules, such as nucleic acids, proteins and polysaccharides. To gain their complete functional activities these biomolecules have to associate with the cellular components, such as the nuclear matrix, cytoskeleton or cell/plasma membranes. Topics discussed in this volume 12 include the synthesis of small nuclear RNAs, DNA-activated protein kinase, interactions of water and proteins in cellular functions, heat-shock protein synthesis and the cytoskeleton during early development.
Book Synopsis Dissertation Abstracts International by :
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Book Synopsis Transcription by : Ronald C. Conaway
Download or read book Transcription written by Ronald C. Conaway and published by Raven Press (ID). This book was released on 1994 with total page 600 pages. Available in PDF, EPUB and Kindle. Book excerpt: Presents a coherent account of many productive lines of investigation, organized as a series of mini-reviews that focus on major research areas including studies on the structure and mechanisms of action of bacterial, viral, and eukaryotic RNA polymerases, and the transcription factors that control their activities. Each review provides a brief but up-to-date account of the progress of research in a particular area, a discussion of the major issues and questions driving that research, and a brief description of the evolving approaches and technologies used to address those questions. Annotation copyright by Book News, Inc., Portland, OR
Book Synopsis Transcriptional Regulation by : Steven L. McKnight
Download or read book Transcriptional Regulation written by Steven L. McKnight and published by . This book was released on 1992 with total page 656 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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Book Synopsis Analysis of Cis-acting Elements and Trans-acting Factors Required for the Expression of Chicken and Drosophila SnRNA Genes by : Zulkeflie Bin Zamrod
Download or read book Analysis of Cis-acting Elements and Trans-acting Factors Required for the Expression of Chicken and Drosophila SnRNA Genes written by Zulkeflie Bin Zamrod and published by . This book was released on 1992 with total page 308 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Abstracts, 23rd Annual Meetings , February 13-26, 1994 by :
Download or read book Abstracts, 23rd Annual Meetings , February 13-26, 1994 written by and published by . This book was released on 1994 with total page 336 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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Book Synopsis Structure and Expression of Chicken U2 and U4 Small Nuclear RNA Genes by : Gina Marie Korf
Download or read book Structure and Expression of Chicken U2 and U4 Small Nuclear RNA Genes written by Gina Marie Korf and published by . This book was released on 1989 with total page 172 pages. Available in PDF, EPUB and Kindle. Book excerpt: In chapter one I report studies on the structure of a chicken gene coding for U2 small nuclear RNA. These studies represent the first cloning, sequencing and characterization of a chicken U2 RNA gene. I report that U2 and U1 RNA genes are organized very differently in the chicken genome, yet they share specific sequence homologies at similar locations in their 5'-flanking DNA. The existence of these conserved sequences at similar locations provided an indication that these sequences are important components of the chicken U2 and U1 RNA gene promoters. In chapter two I present data on the genomic organization of two closely linked chicken U4 RNA genes. One of these genes codes for a well characterized U4 RNA sequence known as U4B RNA. The second gene, U4X, encodes a sequence variant of U4 RNA that was previously unknown. I performed genomic reconstruction experiments that demonstrated that the locus containing the two U4 genes is present in only a single copy per haploid chicken genome. Moreover, these are the only two genes in the chicken genome coding for U4 RNA. Subsequently I performed experiments that demonstrated that U4X RNA exists in chicken tissues, thus proving that the U4X RNA gene is a real gene that is expressed in vivo. In chapter three, I analyze the relative accumulation of U4X and U4B RNA in seven adult chicken tissues and three embryonic chicken tissues. I determined that the relative accumulation of the U4X and U4B RNA gene products can vary in both a tissue and stage-specific manner. Since U4 RNA is a component of the spliceosome, the discovery of differential accumulation of the U4X and U4B RNAs may be relevant to the molecular mechanisms involved in selecting developmentally-regulated alternative splice sites in mRNA precursors. In a final experiment, I examined the RNA of several vertebrate species for the presence of a chicken U4X RNA homolog. The results indicated that a U4X RNA homolog does appear in at least one other avian species.
Book Synopsis Proceedings of the National Academy of Sciences of the United States of America by : National Academy of Sciences (U.S.).
Download or read book Proceedings of the National Academy of Sciences of the United States of America written by National Academy of Sciences (U.S.). and published by . This book was released on 2004 with total page 1334 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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