scholarly journals LCR-dependent gene expression in beta-globin YAC transgenics: detailed structural studies validate functional analysis even in the presence of fragmented YACs

1998 ◽  
Vol 7 (13) ◽  
pp. 2079-2088 ◽  
Author(s):  
K. Peterson
Keyword(s):  
Gene Expression ◽  
Functional Analysis ◽  
Structural Studies ◽  
Beta Globin

scholarly journals 5'-flanking sequences mediate butyrate stimulation of embryonic globin gene expression in adult erythroid cells.

1991 ◽  
Vol 11 (9) ◽  
pp. 4690-4697 ◽  
Author(s):  
J G Glauber ◽  
N J Wandersee ◽  
J A Little ◽  
G D Ginder
Keyword(s):  
Gene Expression ◽  
Cell Differentiation ◽  
Globin Gene ◽  
Beta Globin ◽  
Erythroid Cells ◽  
Adult Chicken ◽  
Beta Globin Gene ◽  
Flanking Sequences ◽  
Murine Erythroleukemia ◽  
Globin Gene Expression

A stable transfection assay was used to test the mechanism by which embryonic globin gene transcription is stimulated in adult erythroid cells exposed to butyric acid and its analogs. To test the appropriate expression and inducibility of chicken globin genes in murine erythroleukemia (MEL) cells, an adult chicken beta-globin gene construct was stably transfected. The chicken beta-globin gene was found to be coregulated with the endogenous adult mouse alpha-globin gene following induction of erythroid differentiation of the transfected MEL cells by incubation with either 2% dimethyl sulfoxide (DMSO) or 1 mM sodium butyrate (NaB). In contrast, a stably transfected embryonic chicken beta-type globin gene, rho, was downregulated during DMSO-induced MEL cell differentiation. However, incubation with NaB, which induces MEL cell differentiation, or alpha-amino butyrate, which does not induce differentiation of MEL cells, resulted in markedly increased levels of transcription from the stably transfected rho gene. Analysis of histone modification showed that induction of rho gene expression was not correlated with increased bulk histone acetylation. A region of 5'-flanking sequence extending from -569 to -725 bp upstream of the rho gene cap site was found to be required for both downregulation of rho gene expression during DMSO-induced differentiation and upregulation by treatment with NaB or alpha-amino butyrate. These data are support for a novel mechanism by which butyrate compounds can alter cellular gene expression through specific DNA sequences. The results reported here are also evidence that 5'-flanking sequences are involved in the suppression of embryonic globin gene expression in terminally differentiated adult erythroid cells.

scholarly journals Wnt7b gene expression and functional analysis in the mussel Mytilus coruscus

2016 ◽  
Vol 15 (4) ◽  
Author(s):  
Y.F. Xu ◽  
X. Liang ◽  
Y.R. Chen ◽  
Y.F. Li ◽  
J.L. Yang
Keyword(s):  
Gene Expression ◽  
Functional Analysis ◽  
Mytilus Coruscus

scholarly journals An atlas of Caenorhabditis elegans chemoreceptor expression

2017 ◽  
Author(s):  
Berta Vidal ◽  
Ulkar Aghayeva ◽  
Haosheng Sun ◽  
Chen Wang ◽  
Lori Glenwinkel ◽  
...  
Keyword(s):  
Gene Expression ◽  
Functional Analysis ◽  
Nervous System ◽  
Gene Family ◽  
Sensory Neurons ◽  
Developmental Studies ◽  
Reporter Expression ◽  
Environmental Signals ◽  
Functional Studies ◽  
Wide Range

ABSTRACTOne goal of modern day neuroscience is the establishment of molecular maps that assign unique features to individual neuron types. Such maps provide important starting points for neuron classification, for functional analysis and for developmental studies aimed at defining the molecular mechanisms of neuron identity acquisition and neuron identity diversification. In this resource paper, we describe a nervous system-wide map of the potential expression sites of 244 members of the largest gene family in the C. elegans genome, rhodopsin-like (class A) GPCR chemoreceptors, using classic gfp reporter gene technology. We cover representatives of all sequence families of chemoreceptors GPCRs, some of which were previously entirely uncharacterized. Most reporters are expressed in a very restricted number of cells, often just in single cells. We assign GPCR reporter expression to all but two of the 37 sensory neuron classes of the sex-shared, core nervous system. Some sensory neurons express a very small number of receptors, while others, particularly nociceptive neurons, co-express several dozen GPCR reporter genes. GPCR reporters are also expressed in a wide range of inter- and motorneurons, as well as nonneuronal cells, suggesting that GPCRs may constitute receptors not just for environmental signals, but also for internal cues. We observe only one notable, frequent association of coexpression patterns, namely in one nociceptive amphid (ASH) and two nociceptive phasmid sensory neurons (PHA, PHB). We identified GPCRs with sexually dimorphic expression and several GPCR reporters that are expressed in a left/right asymmetric manner. We identified a substantial degree of GPCR expression plasticity; particularly in the context of the environmentally-induced dauer diapause stage when one third of all tested GPCRs alter the cellular specificity of their expression within and outside the nervous system. Intriguingly, in a number of cases, the dauer-specific alterations of GPCR reporter expression in specific neuron classes are maintained during postdauer life and in some case new patterns are induced post-dauer, demonstrating that GPCR gene expression may serve as traits of life history. Taken together, our resource provides an entry point for functional studies and also offers a host of molecular markers for studying molecular patterning and plasticity of the nervous system.AUTHOR SUMMARYMaps of gene expression patterns in the nervous system provide an important resource for neuron classification, for functional analysis and for developmental studies that ask how different neurons acquire their unique identities. By analyzing transgenic gfp reporter strains, we describe here the expression pattern of 244 putative chemosensory receptor-encoding genes, which constitute the largest gene family in C.elegans. We show that, as expected, chemoreceptor expression is enriched in chemosensory neurons but it is also expressed in a wide range of interneurons, motorneurons, as well as non-neuronal cells, suggesting that putative chemosensory receptors may not just sense environmental signals but also internal cues. We find that each chemoreceptor is expressed in a few neuron types, often just one, but each neuron type can express a large number of chemoreceptors. Interestingly, we uncovered that chemoreceptor expression is remarkably plastic, particularly in the context of the environmentally-induced dauer diapause stage. Taken together, this molecular atlas of chemosensory receptors provides an entry point for functional studies and offers a host of markers for studying neuronal patterning and plasticity.

scholarly journals Functional analysis and purification of a Pen-2 fusion protein for γ-secretase structural studies

2014 ◽  
Vol 131 (1) ◽  
pp. 94-100
Author(s):  
Oliver Holmes ◽  
Swetha Paturi ◽  
Michael S. Wolfe ◽  
Dennis J. Selkoe
Keyword(s):  
Functional Analysis ◽  
Fusion Protein ◽  
Structural Studies

scholarly journals The essential activities of the bacterial sigma factor

2017 ◽  
Vol 63 (2) ◽  
pp. 89-99 ◽  
Author(s):  
Maria C. Davis ◽  
Christopher A. Kesthely ◽  
Emily A. Franklin ◽  
Shawn R. MacLellan
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Rna Synthesis ◽  
Gene Expression Regulation ◽  
Structure And Function ◽  
Structural Studies ◽  
General Transcription Factors ◽  
Transcription Process ◽  
Promoter Dna ◽  
And Function

Transcription is the first and most heavily regulated step in gene expression. Sigma (σ) factors are general transcription factors that reversibly bind RNA polymerase (RNAP) and mediate transcription of all genes in bacteria. σ Factors play 3 major roles in the RNA synthesis initiation process: they (i) target RNAP holoenzyme to specific promoters, (ii) melt a region of double-stranded promoter DNA and stabilize it as a single-stranded open complex, and (iii) interact with other DNA-binding transcription factors to contribute complexity to gene expression regulation schemes. Recent structural studies have demonstrated that when σ factors bind promoter DNA, they capture 1 or more nucleotides that are flipped out of the helical DNA stack and this stabilizes the promoter open-complex intermediate that is required for the initiation of RNA synthesis. This review describes the structure and function of the σ70 family of σ proteins and the essential roles they play in the transcription process.

scholarly journals Differential gene expression and functional analysis of pit cells from regenerating rat liver

2011 ◽  
Vol 10 (2) ◽  
pp. 678-692 ◽  
Author(s):  
C.S. Xu ◽  
X.G. Chen ◽  
C.F. Chang ◽  
G.P. Wang ◽  
W.B. Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Functional Analysis ◽  
Differential Gene Expression ◽  
Rat Liver ◽  
Regenerating Rat Liver ◽  
Pit Cells ◽  
Differential Gene

scholarly journals Functional Analysis of the Mouse ICER (Inducible cAMP Early Repressor) Promoter: Evidence for a Protein That Blocks Calcium Responsiveness of the CAREs (cAMP Autoregulatory Elements)

1999 ◽  
Vol 13 (7) ◽  
pp. 1207-1217 ◽  
Author(s):  
Darcy A. Krueger ◽  
Dailing Mao ◽  
Elizabeth A. Warner ◽  
Diane R. Dowd
Keyword(s):  
Gene Expression ◽  
Functional Analysis ◽  
Nuclear Protein ◽  
Transcriptional Response ◽  
Binding Activity ◽  
Camp Response Element ◽  
Promoter Sequences ◽  
Inducible Camp Early Repressor ◽  
Relative Binding ◽  
Mutant Promoter

Abstract Although Ca2+ and cAMP mediate their effects through distinct pathways, both signals converge upon the phosphorylation of the cAMP response element (CRE) binding protein, CREB, thereby activating transcription of CRE-regulated genes. In WEHI7.2 thymocytes, cAMP increases the expression of the inducible cAMP early repressor (ICER) gene through CRE-like elements, known as cAMP autoregulatory elements (CAREs). Because Ca2+- and cAMP-mediated transcription converge in WEHI7.2 thymocytes, we examined the effect of Ca2+ fluxes on the expression of the ICER gene in these cells. Despite the presence of multiple CAREs within its promoter, ICER gene transcription was not activated by Ca2+. Moreover, Ca2+ attenuated the stimulatory effect of cAMP on ICER expression. Transient expression of reporter constructs demonstrated that when these CAREs were placed in a different DNA promoter context, the elements became responsive to Ca2+. Detailed studies using chimeric promoter constructs to map the region responsible for blocking the transcriptional response to Ca2+ indicated that a small portion of the ICER promoter was necessary for the effect. Southwestern blot analysis identified a 83-kDa nuclear protein that bound specifically to that region. The relative binding activity of the factor to the ICER promoter and mutant promoter sequences correlated with an inhibition of Ca2+-activated gene expression in WEHI7.2 cells. These data suggest that the factor functions as a putative Ca2+-activated repressor of CREB/CRE-mediated transcription. Thus, depending on the surrounding context in which the CRE is located, CREs of individual genes can be regulated separately by Ca2+ and cAMP despite the convergence of these two signaling pathways.

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