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Biomedical subjects

H Cho

Publications and source records attributed to H Cho.

At least 127 records · Page 7Linked to original sources

A human RNA polymerase II complex associated with SRB and DNA-repair proteins.

We report here the isolation of a human RNA polymerase II complex containing a subset of the basal transcription factors and the human homologues of the yeast SRB (for suppressors of RNA polymerase B) proteins. The complex contains transcriptional coactivators and increases the activation of transcription. In addition, some components of the RNA polymerase II complex participate in DNA repair.

Chromatography, Affinity↗

Applications of tritium NMR to macromolecules: a study of two nucleic acid molecules.

We have tritium labeled two nucleic acid molecules, an 8 kDa DNA oligomer and a 20 kDa 'hammer-head' RNA for tritium NMR investigations. The DNA sequence studied has been previously used in homonuclear studies of DNA-bound water molecules and tritium NMR was expected to facilitate these investigations by eliminating the need to suppress the water resonance in tritium-detected 3H-1H NOESY experiments. We observed the anticipated through-space interactions found in B-form DNA in the NOESY experiments and an unexpected 'antiphase' cross-peak at the water frequency. T1 measurements on the tritiated DNA molecule indicated that relaxation rates were also accelerated for tritium and protons. Tritium NMR spectra of the hammerhead RNA molecule indicated conformational dynamics in the conserved region of the molecule in the absence of Mg2+ and spermine, two components necessary for cleavage. The dynamics were also investigated by 15N-correlated 1H spectroscopy and persisted after the addition of Mg2+ and spermine.

Base Sequence↗

The genetic and molecular dissection of a prototypic circadian system.

A great deal is known about this archetypal circadian system, and it is likely that Neurospora will represent the first circadian system in which it will be possible to provide a complete description of the flow of information from the photoreceptor, through the components of oscillator, out to a terminal aspect of regulation. In Neurospora the strongest case has been made for there being a state variable of clock identified (Hall, 1995), it has now been shown that light resetting of the clock is mediated by the rapid light induction of the gene encoding this state variable, and a number of defined clock-regulated output genes have been identified, in two of which the clock-specific parts of the promoters have been localized. In addition to the importance of these factoids themselves, our efforts towards understanding of this system has allowed the development of tools and paradigms (e.g. Loros et al., 1989; Loros and Dunlap, 1991; Aronson et al., 1994a) that will help to pave the way for proving the identity of clock components in more complex systems, for understanding how clocks are regulated by entraining factors, and for showing how time information eventually is used to regulate the behaviors of clock cells, and of whole organisms.

Animals↗

B cell antigen receptor signaling links biochemical changes in the class II peptide-loading compartment to enhanced processing.

In B cells, processing of antigens in the context of MHC class II molecules is initiated by the binding of antigen to the B cell antigen receptor (BCR). BCR-mediated processing is highly efficient, as a consequence of the BCR's linked roles of delivering antigen to the class II peptide-loading compartment and of signaling for increased antigen-processing activity. Evidence is emerging that receptor signaling regulates intracellular transport through the activities of kinases. These in turn have been implicated in the regulation of small mol. wt GTPases which govern membrane transport. Therefore, we investigated the changes in the phosphoprotein and GTPase profiles associated with the class II peptide-loading compartment following BCR cross-linking. We first show that protein kinase inhibitors, known to block BCR signal transduction, inhibit BCR-enhanced antigen processing, demonstrating the critical dependence of enhanced processing on the signaling activity of the BCR. Consistent with this observation, the phosphoprotein profile of the class II peptide-loading compartment underwent rapid and transient changes following BCR cross-linking. We also observed a marked increase in the low mol. wt GTPases associated with the class II peptide-loading compartment within 5 min of BCR cross-linking. The observed changes in both the phosphoprotein and GTPase profiles associated with the peptide-loading compartment were blocked by kinase inhibitors and were not accompanied by overall gross changes in the protein composition of the subcellular compartments. Thus, signal cascades initiated by BCR cross-linking at the plasma membrane are translated into changes in specific subsets of regulatory proteins associated with the peptide-loading compartment.

Animals↗

Latchkey children.

Children who are regularly left without adult supervision during a significant portion of the day, referred to as 'latchkey children', are a growing social phenomenon. The main reason for the rising prevalence of latchkey children is the increase in dual income and single parent families. Studies on the effects of the latchkey phenomenon report conflicting results. The potential positive consequences include learning to be independent and responsible. The potential negative consequences include loneliness, boredom, fear, academic under-achievement, drug and alcohol abuse, accidental injury, and impairment of the parent-child relationship. Such wide variations in reported consequences in latchkey children might reflect differences in the maturity of the children and in the parent-child relationships prior to entering the latchkey arrangement. Counselling parents about the problems associated with a latchkey arrangement, referring children to an after-school programme, and teaching children self-help skills might minimise the possibility of negative consequences.

Adolescent↗

Uterine endometrial stromal sarcoma with rhabdoid and smooth muscle differentiation.

Uterine and extrauterine tumors composed of cells featuring endometrial stromal cells often show ovarian sex cord-like structures and smooth muscle differentiation. A few cases of endometrial stromal tumors showing rhabdoid differentiation have been reported. The present case is a 20-year-old woman with endometrial stromal sarcoma that had sex cord-like structures, smooth muscle components and rhabdoid differentiation.

Adult↗

Chest pain in children.

Chest pain is usually a benign symptom in children. The most common identifiable causes are musculoskeletal. Often, no cause can be identified. Cardiac disorders are uncommon causes of chest pain children. Most causes can be diagnosed from history and physical examination. Treatment should be directed at the underlying cause. For idiopathic chest pain, reassurance and regular follow-up examinations are important.

Adolescent↗

Refined solution structure and dynamics of the DNA-binding domain of the heat shock factor from Kluyveromyces lactis.

The solution structure of the 92 residue (11 kDa) winged helix-turn-helix DNA-binding domain from the kluyveromyces lactis heat shock factor was refined using a total of 932 NOE, 35 phi, 25 chi 1, 5 chi 2 and 44 hydrogen bond restraints. The overall root-mean-square deviation for structured regions was 0.75(+/- 0.15) A. The three-helix bundle and four-stranded beta-sheet are well defined with rmsd of 0.53(+/- 0.10) A and 0.60(+/- 0.17) A, respectively. Helix H2 is underwound and bent near Pro45. The angle between helix H2 and the proposed recognition helix H3 is 96(+/- 6) degrees. Detailed comparisons are made with the X-ray structure of this protein as well as other structural studies on HSF. Overall, the results are consistent with the earlier studies. Differences are related to protein-protein interactions in the crystal and dynamics in solution. Backbone dynamics was investigated via 15N relaxation. The average R1, R2 and NOE values for residues in segments of secondary structure were 1.9(+/- 0.9) s-1, 7.8(+/- 0.9) s-1 and 0.81(+/- 0.05), respectively. The correlation time based on these data was 5.6(+/- 0.4) ns. Motional order parameters were calculated by fitting the relaxation data to one of three models. Low-order parameters were found for residues that comprise the turn between helices H2 and H3 (residues Lys49 to Phe53), and most strikingly, the 16 residue wing (residues Val68 to Arg83). These data are consistent with the lack of long-range NOEs identified in these regions. The data provide a basis for comparison with results of the protein-DNA complex. The relationship between structure and function is discussed.

Binding Sites↗

Entry of B cell antigen receptor and antigen into class II peptide-loading compartment is independent of receptor cross-linking.

The processing and presentation of Ag by B lymphocytes are initiated by Ag binding to the B cell Ag receptor (BCR). Using subcellular fractionation, we recently identified a compartment in B cells in which functional, processed Ag-class II complexes are formed following BCR-mediated Ag internalization, referred to as the peptide-loading compartment. These studies, however, did not address the transport of Ag or BCR from the cell surface to the peptide-loading compartment. In this work, we describe the intracellular trafficking of Ag and surface Ig (sIg) in B cells and evaluate the effect of cross-linking sIg on this intracellular movement. We show that sIg constitutively transports Ag from the plasma membrane, through endosomes, to the MHC class II peptide-loading compartment. The cross-linking of the BCR increases the rate of internalization of sIg and bound Ag, but does not alter the trafficking pathway. Thus, the delivery of Ag to the class II peptide-loading compartment by the sIg is independent of BCR cross-linking, but can be influenced by BCR cross-linking.

Animals↗

Intracellular transport of invariant chain-MHC class II complexes to the peptide-loading compartment.

Th cells recognize peptide fragments of foreign Ags bound to MHC class II molecules. Upon synthesis in the endoplasmic reticulum, the alpha- and beta-chains of the class II molecules rapidly associate with invariant chains (li). The dissociation of li from class II molecules precedes binding of processed Ag and the formation of SDS-stable alpha beta dimers. We previously showed that functional, processed Ag-class II complexes are assembled in a dense lysosome-like compartment that contains stable class II molecules, but no li, referred to in this work as the peptide-loading compartment. We also identified a separate compartment that contains predominantly SDS-unstable li-class II complexes. Because we were unable to identify known organelle markers associated with this compartment, we refer to it as the X compartment. In this work, we provide results that indicate that the X compartment is composed of transport vesicles that move li-class II complexes to the peptide-loading compartment, where all events in the assembly of processed Ag-class II complexes occur.

Antigens, Differentiation, B-Lymphocyte↗

Defective export of a periplasmic enzyme disrupts regulation of fatty acid synthesis.

Escherichia coli thioesterase I (TesA) encoded by the tesA gene is located in the cellular periplasm. The tesA gene was modified by deletion of the leader sequence such that the mature enzyme was instead localized to the cellular cytosol. Production of thioesterase I in the cytosol results in striking changes in the pattern of E. coli lipid synthesis. In contrast to normal E. coli cells, cells producing cytosolic TesA synthesize large amounts of free fatty acid at all stages of growth. Moreover, cultures of the cytosolic TesA-producing strain continue lipid synthesis (as free fatty acid) in stationary phase whereas lipid synthesis is normally strongly inhibited in such cultures. Surprisingly, production of cytosolic thioesterase I gave only modest inhibition of membrane phospholipid synthesis. These results demonstrate that internalization of a normally secreted enzyme can disrupt normal cellular regulatory mechanisms.

Base Sequence↗

Acute increase of GABAergic neurotransmission exerts a stimulatory effect on GnRH gene expression in the preoptic/anterior hypothalamic area of ovariectomized, estrogen- and progesterone-treated adult female rats.

Although gamma-aminobutyric acid (GABA) is known to play an important role in the regulation of GnRH release from the hypothalamus, GABAergic action on hypothalamic GnRH gene expression is poorly understood. The present study aims to evaluate the effects of several GABAergic compounds on GnRH mRNA and serum LH levels at the times of LH surge induced by estrogen plus progesterone treatment in long-term ovariectomized adult rats. Animals received either aminooxyacetic acid (AOAA, an inhibitor of GABA catabolism, i.p.), muscimol (GABA-A type agonist, i.c.v.) or baclofen (GABA-B type agonist, i.c.v.) 2 h prior to sacrifice. GnRH mRNA in the preoptic/anterior hypothalamic area and serum LH levels were determined by Northern blot analysis and LH radioimmunoassay, respectively. All of three GABA mimetics blocked the LH surge induced by estrogen plus progesterone in a dose-dependent manner. However, inhibition of GABA catabolism with AOAA in a dose range of 10-100 mg/kg b.w. increased GnRH mRNA level by 30%. Activation of GABA-A receptor with muscimol at a low dose (5 nmol) but not at high doses (10 and 30 nmol) elevated GnRH mRNA levels by 60% over the control value. Activation of GABA-B receptor with baclofen augmented GnRH mRNA levels in a dose-dependent manner. These observations indicate that acute increase of GABAergic neurotransmission may differentially regulate the release and GnRH gene expression depending on its receptor subtypes.

Aminooxyacetic Acid↗

Nonnasopharyngeal lymphoepitheliomas (undifferentiated carcinomas) of the upper aerodigestive tract.

Lymphoepitheliomas are malignant tumors of epithelial origin with various amounts of reactive lymphocytic infiltrate. Although initially described in the nasopharynx (World Health Organization type 3 nasopharyngeal carcinoma), these tumors have been identified in various locations throughout the body. A strong association with Epstein-Barr virus (EBV) infection has been established for the nasopharyngeal type. Outside the nasopharynx, lymphoepitheliomas are exceedingly rare in the upper aerodigestive tract, with only isolated case reports of tumors in the larynx, trachea, and hypopharynx. This article features a rare case of lymphoepithelioma of the pyriform sinus. Furthermore, serologic testing, as well as in situ tumor DNA amplification (using the polymerase chain reaction) and hybridization techniques, demonstrated an association of this lesion with EBV infection. The characteristic histopathologic features common to this disease entity are presented, and the literature is reviewed with regard to lymphoepitheliomas of the upper aerodigestive tract outside the nasopharynx. Association of lymphoepitheliomas with EBV infection will be discussed.

Adult↗

Infralabyrinthine approach to vestibular neurectomy in Menière's disease.

The infralabyrinthine approach to vestibular neurectomy was performed in 9 patients with unilateral Menière's disease. According to the AAOO (1972) criteria, 7 of 9 cases were graded as class B and the remaining 2 cases as class C. Otherwise, according to the AAO-HNS (1985) criteria, 6 patients who could be followed over 2 years were all graded as 'complete' at the vertigo control. The compensation of the spontaneous vestibular signs was rapid in the first 2 postoperative weeks, though an occasional imbalance on movement persisted even 3 years after the operation. No specific caloric reaction was elicited in any patient after warm or cold water irrigation of the operated side in any postoperative period. There have been no serious complications except a delayed facial palsy that appeared in one case one week after surgery. This approach offers access to the vestibular nerve with minimal risk and morbidity.

Adult↗

The intracellular assembly of antigenic-peptide-class II complexes.

The immune system employs remarkable strategies to ensure that foreign antigens, from the most complex pathogens to the simplest proteins, are displayed on the surfaces of cells which are targets of T lymphocyte recognition. At the heart of these strategies is the molecular transformation of a soluble protein antigen to a complex of a small peptide containing the antigenic determinant bound to a cell surface Major Histocompatibility Complex class I or class II protein. This process is termed antigen presentation. Progress in a variety of laboratories over the last several years has yielded a wealth of information about the molecular mechanisms underlying antigen presentation, providing potential new approaches to vaccine design. Here we describe recent studies in our laboratory aimed at elucidating the intracellular site in B lymphocytes in which antigenic peptide-class II complexes are assembled for recognition by helper T cells and the regulation of this assembly process. Our results suggest that processed antigen-class II complexes are assembled in a unique compartment in the endocytic route which contains all the necessary cellular and molecular machinery for assembly and that B cells regulate the assembly process in response to external and internal signals.

Antigen-Presenting Cells↗