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At least 1,423 records · Page 79Linked to original sources
PubMed1994

Primary structure and functional expression of a guinea pig kappa opioid (dynorphin) receptor.

A full-length cDNA encoding the guinea pig kappa opioid (dynorphin) receptor has been isolated. The deduced protein contains 380 aa and seven hydrophobic alpha-helices characteristic of the G protein-coupled receptors. This receptor is 90% identical to the mouse and rat kappa receptors, with the greatest level of divergence in the N-terminal region. When expressed in COS-7 cells, the receptor displays high affinity and stereospecificity toward dynorphin peptides and other kappa-selective opioid ligands such as U50, 488. It does not bind the mu- and delta-selective opioid ligands. The expressed receptor is functionally coupled to G protein(s) to inhibit adenylyl cyclase and Ca2+ channels. The guinea pig kappa receptor mRNA is expressed in many brain areas, including the cerebellum, a pattern that agrees well with autoradiographic maps of classical guinea pig kappa binding sites. Species differences in the pharmacology and mRNA distribution between the cloned guinea pig and rat kappa receptors may be worthy of further examination.

Amino Acid Sequence↗
PubMed2003

N-terminal extensions of the human AMPD2 polypeptide influence ATP regulation of isoform L.

Human tissues and cells express three AMP deaminase (AMPD) isoforms containing divergent N-terminal domains, and each member of the multigene family encoding these enzymes produces alternative transcripts that confer additional N-terminal divergence through extensions and cassette-type substitutions. Available data suggest that divergent N-terminal domains can influence AMPD isoform behavior, but the functional significance for additional divergence within each enzyme is unknown. Three isoform L (AMPD2) variants, 1A/2, 1B/2, and 1B/3, contain N-terminal extensions of 47, 128, and 53 amino acids, respectively. This study has determined the kinetic and regulatory behaviors of these three isoform L enzymes in the presence of positive (ATP) and negative (phosphate) allosteric effectors. All display nearly identical kinetic parameters and regulatory responses in the presence of phosphate alone, or in combination with ATP. Regulation by ATP is biphasic and the three isoform L enzymes also exhibit similar activation profiles and maximum initial velocities at 2-3mM in the presence of 1mM phosphate, whereas higher concentrations of phosphate suppress this activation. However, maximum initial velocities are achieved at lower ATP concentrations (0.8-1.5mM) in the absence of phosphate and under these conditions 1B/2 is less active, 1B/3 is more active, and 1A/2 is similarly active when compared to 1mM phosphate over the range of ATP concentrations found in non-muscle cells (0.8-3.7mM). These combined results suggest that isoform L enzymes are designed to function under different metabolic conditions encountered in the non-striated muscle environments where they are expressed.

AMP Deaminase↗
PubMed1999

Regulated expression of a highly conserved regulatory gene cluster is necessary for controlling photosynthesis gene expression in response to anaerobiosis in Rhodobacter capsulatus.

We utilized primer extension analysis to demonstrate that the divergently transcribed regB and senC-regA-hvrA transcripts contain stable 5' ends 43 nucleotides apart within the regB-senC intergenic region. DNA sequence analysis indicates that this region contains two divergent promoters with overlapping sigma70 type -35 and -10 promoter recognition sequences. In vivo analysis of expression patterns of regB::lacZ and senC-regA-hvrA::lacZ reporter gene fusions demonstrates that the regB and senC-regA-hvrA transcripts are both negatively regulated by the phosphorylated form of the global response regulator RegA. DNase I protection assays with a constitutively active variant of RegA indicate that RegA binds between regB and senC overlapping -10 and -35 promoter recognition sequences. Two mutations were also isolated in a regB-deficient background that increased expression of the senC-regA-hvrA operon 10- and 5-fold, respectively. As a consequence of increased RegA expression, these mutants exhibited elevated aerobic and anaerobic photosynthesis (puf) gene expression, even in the absence of the sensor kinase RegB. These results indicate that autoregulation by RegA is a factor contributing to the maintenance of an optimal low level of RegA expression that allows responsiveness to activation by phosphorylation.

Anaerobiosis↗
PubMed1994

Genetic control of early embryogenesis in the red flour beetle, Tribolium castaneum.

The power of genetic analysis possible with the fruit fly, Drosophila melanogaster, has yielded a detailed understanding of pattern formation controlled by homeotic and segmentation genes in early embryogenesis. We are studying the genetic regulation of embryogenesis in the red flour beetle, Tribolium castaneum. The dynamic process of germ rudiment formation and sequential segmentation exhibited by Tribolium provides a context different than Drosophila within which to assess the function of homeotic and segmentation gene homologs. Our analyses of the genes in the HOM-C suggest many similarities in structure and function with the well-characterized Drosophila genes. Abdominal resembles its Drosophila homolog abdominal-A in functioning to establish segmental identities in the abdomen, such that in each case mutations result in homeotic transformations to PS6. Although the anterior functional boundary of abdominal-A homologs is precisely conserved, the domain within which Abdominal is important extends more posterior than that of abdominal-A. The final expression pattern of the segmentation gene engrailed in Tribolium is identical to Drosophila, suggesting that these homologs are involved in a conserved developmental process. However, as expected the development of that pattern is different; engrailed stripes anticipate the formation of each new segment as they appear sequentially in the elongating germ band. Although the grasshopper even-skipped and fushi tarazu homologs are not apparently important in segmentation, the expression patterns of the Tribolium homologs strongly suggest that they have gained a role in segmentation in the lineage leading to beetles and flies. Nevertheless, differences between Tribolium and Drosophila in the dynamics of even-skipped expression and the fushi tarazu mutant phenotype indicate divergence in the regulation and roles of these genes.

Animals↗
PubMed2003

Expressing functional siRNAs in mammalian cells using convergent transcription.

BACKGROUND: The use of small interfering RNAs (siRNAs) as genetic inhibitors of gene expression has been shown to be an effective way of studying gene function in mammalian cells. Recently, different DNA vectors for expression of small hairpin RNAs (shRNAs) or co-expression of sense and antisense RNAs have been developed that direct siRNA-mediated gene silencing. One expression cassette design that has been used to express long sense and antisense RNAs in non-mammalian cell types is symmetric transcription using convergent promoters. However, convergent transcription as a way to generate functional siRNAs in mammalian cells has not been reported. This vector design permits the generation of expression constructs containing no repeat sequences, but capable of inducing RNA interference (RNAi)-mediated gene silencing. RESULTS: With the aim of simplifying the construction of RNAi expression vectors, we report on the production and application of a novel convergent promoter cassette capable of expressing sense and antisense RNAs, that form double-stranded RNA, and mediate gene silencing in mammalian cells. We use this cassette to inhibit the expression of both the EGFP transgene and the endogenous TP53 gene. The gene silencing effect is Dicer-dependent and the level of gene inactivation achieved is comparable to that produced with synthetic siRNA. Furthermore, this expression system can be used for both short and long-term control of specific gene expression in mammalian cells. CONCLUSION: The experiments performed in this study demonstrate that convergent transcription can be used in mammalian cells to invoke gene-specific silencing via RNAi. This method provides an alternative to expression of shRNAs and co-expression of sense and antisense RNAs from independent cassettes or a divergent promoter. The main advantage of the present vector design is the potential to produce a functional siRNA expression cassette with no repeat sequences. Furthermore, the cassette design reported is ideal for both routine use in controlling specific gene expression and construction of randomised RNAi expression libraries for use in unbiased forward genetic selections.

Cell Line↗
PubMed2026

Genome-wide identification of HCT gene family in sugarcane (Saccharum spp. hybrid) and characterization of putative cis-elements in gene regulation.

BACKGROUND: Sugarcane (Saccharum spp. hybrid) is a globally important crop, and its bagasse can be converted into bioethanol and other industrial products. Lignin, a core component of sugarcane cell walls, plays a crucial role in bagasse quality and lodging resistance. Shikimic acid hydroxycinnamyl transferase (HCT) is the key enzyme in lignin biosynthesis. However, the HCT gene family in sugarcane and its regulatory roles in sugarcane remain poorly understood. RESULTS: A total of 663 HCT genes (including alleles) were identified in the Saccharum hybrid R570 genome, which were classified into six groups (I-VI) and were unevenly distributed across 77 chromosomes. Bioinformatics analysis revealed that the subgroups of R570HCTs had similar gene structures, suggesting conserved functions. Moreover, the different subgroups presented unique putative cis‑element distribution patterns. Transcriptome data indicated that some R570HCTs exhibited significant spatiotemporal and tissue‑specific expression patterns. Further Pearson correlation analysis between putative cis‑element distribution and normalized expression values at the subgroup level revealed that light-responsive elements (L‑box and GA‑motif) were positively correlated with R570HCT expression, and different subgroups formed a complex regulatory network by integrating hormone response and stress elements. Importantly, this subgroup-level correlation was cross-validated by comparing the cis‑element clustering heatmap with the expression heatmap, revealing consistent enrichment patterns. CONCLUSIONS: The study's findings provide novel insights into the correlation among motifs, putative cis‑elements, and gene expression, and propose a cross-validated framework for understanding regulatory divergence among HCT subfamilies in polyploid sugarcane, serving as a hypothesis generating resource for future research on R570HCT expression.

Saccharum↗
PubMed2005

Influence of gap and overlap paradigms on saccade latencies and vergence eye movements in seven-year-old children.

The latency of eye movements is influenced by the fixation task; when the fixation stimulus is switched off before the target presentation (gap paradigm) the latency becomes short and express movements occur. In contrast, when the fixation stimulus remains on when the target appears (overlap paradigm), eye movement latency is longer. Several previous studies have shown increased rates of express saccades in children; however the presence of an express type of latency for vergence and combined movements in children has never been explored. The present study examines the effects of the gap and the overlap paradigms on horizontal saccades at far (150 cm) and at close (20 cm) viewing distances, on vergence along the median plane, and on saccades combined with convergence or divergence in 15 normal seven-year-old children. The results show that the gap paradigm produced shorter latency for all eye movements than the overlap paradigm, but the difference was only significant for saccades at close viewing distances, for divergence (pure and combined), and for saccades combined with vergence. The gap paradigm produced significantly higher rates of express latencies for saccades at close viewing distances, for divergence, and for saccades combined with divergence; in contrast, the frequencies of express latencies for saccades at far viewing distances and for convergence (pure or combined) were similar in the gap and the overlap paradigms. Interestingly, the rate of anticipatory latencies (<80 ms) was particularly high for divergence in the gap paradigm. Our collective findings suggest that the initiation of saccades at close viewing distances and of divergence is more reflexive, particularly in the gap paradigm. The finding of frequent anticipatory divergence that occurs at similar rates for seven-year-old children (this study) and for adults (Coubard et al., 2004, Exp Brain Res 154:368-381) indicates that predictive initiation of divergence is dominant.

Aging↗
PubMed1990

Two Drosophila beta tubulin isoforms are not functionally equivalent.

We have tested the functional capacity of different beta tubulin isoforms in vivo by expressing beta 3-tubulin either in place of or in addition to beta 2-tubulin in the male germ line of Drosophila melanogaster. The testes-specific isoform, beta 2, is conserved relative to major metazoan beta tubulins, while the developmentally regulated isoform, beta 3, is considerably divergent in sequence. beta 3-tubulin is normally expressed in discrete subsets of cells at specific times during development, but is not expressed in the male germ line. beta 2-Tubulin is normally expressed only in the postmitotic germ cells of the testis, and is required for all microtubule-based functions in these cells. The normal functions of beta 2-tubulin include assembly of meiotic spindles, axonemes, and at least two classes of cytoplasmic microtubules, including those associated with the differentiating mitochondrial derivatives. A hybrid gene was constructed in which 5' sequences from the beta 2 gene were joined to protein coding and 3' sequences of the beta 3 gene. Drosophila transformed with the hybrid gene express beta 3-tubulin in the postmitotic male germ cells. When expressed in the absence of the normal testis isoform, beta 3-tubulin supports assembly of one class of functional cytoplasmic microtubules. In such males the microtubules associated with the membranes of the mitochondrial derivatives are assembled and normal mitochondrial derivative elongation occurs, but axoneme assembly and other microtubule-mediated processes, including meiosis and nuclear shaping, do not occur. These data show that beta 3 tubulin can support only a subset of the multiple functions normally performed by beta 2, and also suggest that the microtubules associated with the mitochondrial derivatives mediate their elongation. When beta 3 is coexpressed in the male germ line with beta 2, at any level, spindles and all classes of cytoplasmic microtubules are assembled and function normally. However, when beta 3-tubulin exceeds 20% of the total testis beta tubulin pool, it acts in a dominant way to disrupt normal axoneme assembly. In the axonemes assembled in such males, the doublet tubules acquire some of the morphological characteristics of the singlet microtubules of the central pair and accessory tubules. These data therefore unambiguously demonstrate that the Drosophila beta tubulin isoforms beta 2 and beta 3 are not equivalent in intrinsic functional capacity, and furthermore show that assembly of the doublet tubules of the axoneme imposes different constraints on beta tubulin function than does assembly of singlet microtubules.

Animals↗
PubMed1999

A unified paraxial approach to astigmatic optics.

In Gaussian optics properties such as dioptric power, lateral and angular magnification and thickness are simple scalar concepts. In linear optics, the optics of thick astigmatic systems, however, these concepts generalize to three-dimensional concepts in some cases (the dioptric power of thin systems, for example) and to four-dimensional concepts in general. As a result, the quantitative treatment of these properties in astigmatic systems presents challenges to the researcher in optometry, ophthalmology, and vision science. Considerable progress has been made only in the case of dioptric power. This paper presents a generalized approach to astigmatic optics which allows different physical properties to be treated in the same way: the theory is unified and, in a sense, complete. Mathematical and statistical methods developed for treating one concept become directly applicable to others. The paraxial optical properties of any optical system are completely defined by the 4 x 4 ray transfer matrix, called here the (ray) transference. The transference defines four fundamental properties of an optical system, tentatively called here positional magnification, optical thickness, divergence, and directional magnification. They are the four 2 x 2 submatrices A, B, C, and D of the transference. Each fundamental property is a modification of a familiar concept. Divergence is the negative of dioptric power expressed as the dioptric power matrix F. The four fundamental optical properties A, B, C, and D, and the derived property F, despite being different physically, all have the same underlying mathematical structure. This fact is exploited in developing a unified theory. The theory is complete in the sense that the fundamental properties fully characterize the paraxial optics of any system. The paper presents a general treatment that applies to any of the five properties. The implications are far reaching and extend beyond what can be described in the paper. Dioptric power of thin systems is treated as a particular application of the general theory. The result is the resolution of a number of issues of current interest to the researcher. It is shown, for example, that root-mean-squared (curvital) power, root-mean-squared torsional power, and length of the power vector (or dioptric strength) have a Pythagorean relationship, the power vector being the hypotenuse. Mean-squared curvital and torsional powers are in effect the area enclosed by polar profiles of curvital and torsional power, respectively. The full character of dioptric power cannot be represented by a single vector in the usual sense of the term. Two vectors are required: they are the meridional (vector) power and the orthogonal (vector) power, both of which are associated with the reference meridian. The power along a meridian (often thought of as a scalar or as two scalars) is a vector, the meridional power. This meridional power has components along (the meridional component of the meridional power) and perpendicular to (the orthogonal component of the meridional power) the meridian. In the literature, these components are the curvital power and the negative of the torsional power, respectively. The paper also examines the generalization of these results to the dioptric power of thick systems. Dioptric power is not a fundamental optical property but a derived property. Divergence, the negative of dioptric power, is the corresponding fundamental property. The theory described here is ray-based. The concept of the wavefront is unnecessary. The many formulas and concepts that apply in the context of dioptric power apply directly to the fundamental properties as well. The theory has the potential to provide a complete framework for future studies of astigmatic systems and could systematize the approach to and enhance the knowledge of astigmatism.

Astigmatism↗
PubMed2002

Prosodic characteristics of skilled reading: fluency and expressiveness in 8-10-year-old readers.

Statistical methods of describing prosody were used to study fluency, expressiveness and their relationship among 8-10-year-old readers. 67 children were rated on fluency and expressiveness. The two were partially independent in the full sample: expressiveness rarely occurred without fluency, but fluency occurred without expressiveness. A balanced subsample of 24 was selected for closer instrumental and statistical analysis. There were robust relationships between fluency and measures associated with temporal organization.between expressiveness and variables associated with pitch mobility; and Interactions indicated that the relationships were not simple. Differences between groups depended on sentence content and position-expressive readers distinguished sentences more sharply according to content, and the groups diverged on some measures as the passage progressed. Also, measures associated primarily with either fluency or expression often showed secondary sensitivity to the other: temporal organization was associated with fluency, but worsened over time among inexpressive readers; and readers who were both fluent and expressive were distinctive in several respects. Some measures offer a basis for rules aimed at assigning individuals to skill categories, particularly the magnitude of pitch movements and reading time per syllable. The rules distinguish well among readers who were either at one of the extremes of skill, or fluent but inexpressive; it is harder to discriminate among the other readers (who have mixed skill patterns). The effects suggest psychological hypotheses about the underlying mechanisms.

Child↗
PubMed2025

Analysis of duplication and possible sub-functionalization of wing gene network components in pea aphids.

A fundamental focus of evolutionary-developmental biology is uncovering the genetic mechanisms responsible for the gain and loss of characters. One approach to this question is to investigate changes in the coordinated expression of a group of genes important for the development of a character of interest (a gene regulatory network). Here we consider the possibility that modifications to the wing gene regulatory network (wGRN), as defined by work primarily done in Drosophila melanogaster, were involved in the evolution of wing dimorphisms of the pea aphid (Acyrthosiphon pisum). We hypothesize that this may have occurred via changes in expression levels or duplication followed by sub-functionalization of wGRN components. To test this, we annotated members of the wGRN in the pea aphid genome and assessed their expression levels in first and third nymphal instars of winged and wingless morphs of males and asexual females. We find that only two of the 32 assessed genes exhibit morph-biased expression. We also find that three wing genes (apterous (ap), warts (wts), and decapentaplegic (dpp)) have undergone gene duplication. In each case, the resulting paralogs show signs of functional divergence, exhibiting either sex-, morph-, or stage-specific expression. Two gene duplicates, wts2 and dpp3, are of particular interest with respect to wing dimorphism, as they exhibit a wingless male-specific isoform and wingless male-biased expression, respectively. These results supplement our understanding of trends in developmental gene network evolution, such as side-stepping pleiotropic constraint via duplication and sub-functionalization, underlying the emergence of novel phenotypes.

dimorphism↗
PubMed2005

EMBRYONIC FACTOR 1 encodes an AMP deaminase and is essential for the zygote to embryo transition in Arabidopsis.

Fusion of the egg and the sperm cells in plants produces a zygote that develops into an embryo. Screening of ethyl methanesulfonate-mutagenized populations of Arabidopsis led to the identification of EMBRYONIC FACTOR 1 (FAC1), a locus that gives a zygote-lethal phenotype when mutated. The FAC1 gene was identified by positional cloning and confirmed by a genetic complementation test against a T-DNA insertion allele. It encodes an AMP deaminase (AMPD) that is known in human and yeast to convert AMP to IMP to maintain the energy potential. Expression of FAC1 in a yeast AMPD mutant after removal of its N-terminal putative transmembrane domain complemented the mutant phenotype, suggesting a functional conservancy but a structural divergence through evolution. Although a low level of FAC1 expression was observed in all organs tested, using a reporter construct we observed a significantly increased FAC1 expression in the zygote, early embryo and endosperm. Furthermore, during somatic embryogenesis, a high level of FAC1 expression was observed in developing embryos including putative embryogenic cells. FAC1, therefore, represents one of the earliest expressed genes known in plants. It may act through AMP depletion to provide sufficient energy for the zygote to proceed through development.

AMP Deaminase↗
PubMed2005

Modulation of gene expression in precancerous rat esophagus by dietary zinc deficit and replenishment.

Zinc deficiency in rats enhances esophageal cell proliferation, causes alteration in gene expression, and promotes esophageal carcinogenesis. Zinc replenishment rapidly induces apoptosis in the esophageal epithelium thereby reversing cell proliferation and carcinogenesis. To identify zinc-responsive genes responsible for these divergent effects, we did oligonucleotide array-based gene expression profiling analyses in the precancerous zinc-deficient esophagus and in zinc-replenished esophagi after treatment with intragastric zinc compared with zinc-sufficient esophagi. Thirty-three genes (21 up-regulated and 12 down-regulated) showed a > or = 2-fold change in expression in the hyperplastic zinc-deficient versus zinc-sufficient esophageal epithelia. Expression of genes involved in cell division, survival, adhesion, and tumorigenesis were markedly changed. The zinc-sensitive gene metallothionein-1 (MT-1 was up-regulated 7-fold, the opposite of results for small intestine and liver under zinc-deficient conditions. Keratin 14 (KRT14, a biomarker in esophageal tumorigenesis), carbonic anhydrase II (CAII, a regulator of acid-base homeostasis), and cyclin B were up-regulated >4-fold. Immunohistochemistry showed that metallothionein and keratin 14 proteins were overexpressed in zinc-deficient esophagus, as well as in lingual and esophageal squamous cell carcinoma from carcinogen-treated rats, emphasizing their roles in carcinogenesis. Calponin 1 (CNN1, an actin cross-linking regulator) was down-regulated 0.2-fold. Within hours after oral zinc treatment, the abnormal expression of 29 of 33 genes returned to near zinc-sufficient levels, accompanied by reversal of the precancerous phenotype. Thus, we have identified new molecular markers in precancerous esophagus and showed their restoration by zinc replenishment, providing insights into the interaction between zinc and gene expression in esophageal cancer development and prevention.

4-Nitroquinoline-1-oxide↗
PubMed1996

Pseudogenes in ribonuclease evolution: a source of new biomacromolecular function?

Bovine seminal ribonuclease (RNase) diverged from pancreatic RNase after a gene duplication ca. 35 million years ago. Members of the seminal RNase gene family evidently remained as unexpressed pseudogene for much of its evolutionary history. Between 5 and 10 million years ago, however, after the divergence of kudu but before the divergence of ox, evidence suggests that the pseudogene was repaired and expressed. Intriguingly, detailed analysis of the sequences suggests that the repair may have involved gene conversion, transfer of information from the pancreatic gene to the RNase pseudogene. Further, the ratio of non-silent to silent substitutions suggests that the pancreatic RNases are divergently evolving under functional constraints, the seminal RNase pseudogenes are diverging under no functional constraints, while the genes expressed in the seminal plasma are evolving extremely rapidly in their amino acid sequences, as if to fulfil a new physiological role.

Amino Acid Sequence↗
PubMed2005

Smad7 abrogates transforming growth factor-beta1-mediated growth inhibition in COLO-357 cells through functional inactivation of the retinoblastoma protein.

Smad7 is overexpressed in 50% of human pancreatic cancers. COLO-357 pancreatic cancer cells engineered to overexpress Smad7 are resistant to the actions of transforming growth factor-beta1 (TGF-beta1) with respect to growth inhibition and cisplatin-induced apoptosis but not with respect to modulation of gene expression. To delineate the mechanisms underlying these divergent consequences of Smad7 overexpression, we studied the effects of Smad7 on TGF-beta1-dependent signaling pathways and cell cycle regulating proteins. TGF-beta1 induced the phosphorylation of MAPK, p38 MAPK, and AKT2 irrespective of the levels of Smad7, and inhibitors of these pathways did not alter TGF-beta1 actions on cell growth. By contrast, Smad7 overexpression interfered with TGF-beta1-mediated attenuation of cyclin A and B levels, inhibition of cdc2 dephosphorylation and CDK2 inactivation, up-regulation of p27, and the maintenance of the retinoblastoma protein (RB) in a hypophosphorylated state. Smad7 also suppressed TGF-beta1-mediated inhibition of E2F activity but did not alter TGF-beta1-mediated phosphorylation of Smad2, the nuclear translocation of Smad2/3/4, or DNA binding of the Smad2/3/4 complex. Although Smad7 did not associate with the type I TGF-beta receptor (TbetaRI), SB-431542, an inhibitor of the kinase activity of this receptor, blocked TGF-beta1-mediated effects on Smad-2 phosphorylation. These findings point toward a novel paradigm whereby Smad7 acts to functionally inactivate RB and de-repress E2F without blocking the activation of TbetaRI and the nuclear translocation of Smad2/3, thereby allowing for TGF-beta1 to exert effects in a cancer cell that is resistant to TGF-beta1-mediated growth inhibition.

Active Transport, Cell Nucleus↗
PubMed2003

Signaling effects of demethylasterriquinone B1, a selective insulin receptor modulator.

A possible breakthrough in the treatment of diabetes was made with the discovery that a fungal natural product, demethylasterriquinone B1 (DAQ B1), is an orally active, small-molecule mimic of insulin. Subsequent work has shown that the glucose-lowering effects of DAQ B1 are not accompanied by enhanced vascular proliferation, which is a side effect of chronic insulin administration that can lead to arteriosclerosis. Our recent short and modular total synthesis of DAQ B1 could be readily modified to create congeners and afforded ample supplies of the natural product, which permitted intracellular signal transduction of DAQ B1 to be examined. The activities of DAQ B1 and over a dozen related structures were studied for insulin receptor (IR) and insulin receptor substrate-1 phosphorylation. Examination of the effect of DAQ B1 on kinases downstream of the IR in insulin signal transduction showed selective activation of Akt kinase (a metabolic effect) but not of extracellular-regulated kinase (a proliferative effect). The influence of DAQ B1 on gene expression (determined by a microarray study) was also divergent from that of insulin, which activates both proliferative and metabolic pathways. The action of DAQ B1 as a selective insulin receptor modulator can be accounted for by its ability to selectively activate one kinase among the many emanating from insulin receptor autophosphorylation and its reduced effects on gene expression.

3T3 Cells↗
PubMed1990

Analysis of the expression of murine lambda genes transfected into immunocompetent cell lines.

Hybridoma cell lines were transfected with plasmids containing either a rearranged lambda 1 or a rearranged lambda 2 mouse gene. The levels of lambda-chains synthesized by these transfectants were very low or undetectable. Activation of the expression of the lambda 2 gene was achieved artificially by deleting a portion of the region upstream of the promoter. Analogous deletions in the fragment containing the lambda 1 gene did not result in gene activation suggesting that the upstream sequences of lambda 1 and lambda 2 genes have diverged enough to allow differential regulation of their expression. However, both genes were activated by insertion, at a position upstream of the promoter, of a fragment containing the K-chain gene enhancer. These results suggest that the complete set of sequence elements that mediate lambda gene activation during normal B-cell differentiation are not all contained in the fragments of genomic DNA cloned so far, and thus, at least some of them must be located at a considerable distance from the promoter.

Animals↗
PubMed1996

Cell stress-regulated human major histocompatibility complex class I gene expressed in gastrointestinal epithelium.

Conventional major histocompatibility complex (MHC) class I genes encode molecules that present intracellular peptide antigens to T cells. They are ubiquitously expressed and regulated by interferon gamma. Two highly divergent human MHC class I genes, MICA and MICB, are regulated by promoter heat shock elements similar to those of HSP70 genes. MICA encodes a cell surface glycoprotein, which is not associated with beta 2-microglobulin, is conformationally stable independent of conventional class I peptide ligands, and almost exclusively expressed in gastrointestinal epithelium. Thus, this MHC class I molecule may function as an indicator of cell stress and may be recognized by a subset of gut mucosal T cells in an unusual interaction.

Animals↗