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T Luo

Publications and source records attributed to T Luo.

At least 37 records · Page 2Linked to original sources
PubMed1997

Identification of an autonomously initiating RNA polymerase III holoenzyme containing a novel factor that is selectively inactivated during protein synthesis inhibition.

Transcription by RNA polymerase III (Pol III) requires multiple general initiation factors that, in isolated form, assemble onto the promoter in an ordered fashion. Here, it is shown that all components required for transcription of the VA1 and tRNA genes, including TFIIIB, TFIIIC, and RNA Pol III, can be coimmunopurified from a HeLa cell line that constantly expresses a FLAG epitope-tagged subunit of human RNA Pol III. This finding of an RNA Pol III "holoenzyme" suggests similarities between transcription initiation by RNA Pol II and RNA Pol III and has led to the identification of a novel general initiation factor (TDF, translation dependent factor) that is present within the holoenzyme. TDF is selectively inactivated during protein synthesis inhibition by cycloheximide and at a late stage of adenovirus infection, thus accounting for the loss of RNA Pol III-mediated transcription of the tRNA and VA RNA genes under these conditions. On the basis of these observations, possible mechanisms for the global regulation of transcription by RNA Pol III and for disassembly of RNA Pol III initiation complexes are proposed.

Coenzymes↗
PubMed1997

Bacillus subtilis Pro-sigmaE fusion protein localizes to the forespore septum and fails to be processed when synthesized in the forespore.

Endospore formation in Bacillus subtilis begins with an asymmetric cell division that partitions the bacterium into mother cell and forespore compartments. Mother cell-specific gene expression is initiated by sigmaE, a transcription factor that is active only in the mother cell but which existed as an inactive precursor (pro-sigmaE) in the predivisional cell. Activation of pro-sigmaE involves the removal of 27 amino acids from its amino terminus. A chimera of pro-sigmaE and the green fluorescent protein (GFP) was expressed from either the normal sigE promoter (P(spoIIG)), which places pro-sigmaE::GFP in both mother cell and forespore compartments, or the forespore-specific promoter (P(dacF)), which produces pro-sigmaE::GFP only in the forespore compartment. The pro-sigmaE::GFP expressed from P(spoIIG), but not P(dacF), was converted to a lower-molecular-weight form by a mechanism dependent on gene products (SpoIIGA and sigmaF) that are essential for normal pro-sigmaE processing. This finding is consistent with the pro-sigmaE processing reaction occurring only in the mother cell compartment. In processing-deficient cells, pro-sigmaE::GFP was found to accumulate at the septal membrane, a location where its processing apparatus would be susceptible to triggering from the adjoining forespore.

Bacillus subtilis↗
PubMed1997

Stress activation of Bacillus subtilis sigma B can occur in the absence of the sigma B negative regulator RsbX.

Environmental stress activates sigma B, the general stress response sigma factor of Bacillus subtilis, by a pathway that is negatively controlled by the RsbX protein. To determine whether stress activation of sigma B occurs by a direct effect of stress on RsbX, we constructed B. subtilis strains which synthesized various amounts of RsbX or lacked RsbX entirely and subjected these strains to ethanol stress. Based on the induction of a sigma B-dependent promoter, stress activation of sigma B can occur in the absence of RsbX. Higher levels of RsbX failed to detectably influence stress induction, but reduced levels of RsbX resulted in greater and longer-lived sigma B activation. The data suggest that RsbX is not a direct participant in the sigma B stress induction process but rather serves as a device to limit the magnitude of the stress response.

Bacillus subtilis↗
PubMed1997

Infectivity enhancement by human immunodeficiency virus type 1 Nef is independent of its association with a cellular serine/threonine kinase.

Nef proteins from human immunodeficiency virus type 1 isolate SF2 (HIV-1SF2) and simian immunodeficiency virus isolate mac239 (SIVmac239) have been found to associate with a cellular serine/threonine kinase designated NAK. We have recently shown that the association of Nef with NAK is isolate dependent. To identify the structural basis for Nef-kinase association, several chimeric molecules were constructed between SF2 Nef (binding NAK) and 233 Nef (a primary isolate not binding NAK) and stably expressed in HuT-78 human T cells via retrovirus-mediated gene transfer. The Nef 233/SF2/SF2 chimera in which the N-terminal 37 amino acids of SF2 Nef were replaced by those of 233 Nef showed the same ability as SF2 Nef to bind NAK. The Nef 233/SF2/233 chimera in which the N-terminal 37 amino acids and the C-terminal 72 amino acids of SF2 Nef were replaced by corresponding sequences from 233 Nef completely lost the ability to associate with the kinase activity. Furthermore, replacement of the C-terminal 72 amino acids of 233 Nef with the equivalent SF2 sequence (chimera 233/233/SF2) fully restored kinase association to 233 Nef. These results suggest that (i) the core of Nef is not sufficient for NAK binding, (ii) the C terminus of SF2 Nef contains structural determinants important for association with NAK, and (iii) the failure of 233 Nef to bind NAK is due to a defect in its C terminus. Taking advantage of the C terminus of 233 Nef being nonfunctional and using an infectious clone of HIV-1SF2, we show that association with NAK is not required for Nef-mediated infectivity enhancement. While the strong and reproducible association of some Nef isolates with NAK has been clearly established, the role of NAK in Nef function remains to be fully elucidated.

Cell Line, Transformed↗
PubMed1997

Induction of phosphorylation of human immunodeficiency virus type 1 Nef and enhancement of CD4 downregulation by phorbol myristate acetate.

The nef gene of the human and simian immunodeficiency viruses (HIV and SIV) encodes a 27 to 34 kDa myristoylated protein that induces downregulation of CD4 from the cell surface and enhances virus infectivity. As shown by experiments on SIV-infected adult macaques, Nef is important in pathogenesis and disease progression. In vitro, protein kinase C (PKC) phosphorylates Nef, but the role of phosphorylation in the function and expression of this protein has not yet been determined. Here we show that in HIV type 1-infected cells, phosphorylation of Nef increased 8- to 12-fold after treatment with phorbol myristate acetate and phytohemagglutinin (PMA/PHA). Basal and PMA/PHA-induced phosphorylation occurred on serine residues of Nef and was independent of other HIV proteins. The PMA/PHA-induced phosphorylation of Nef was inhibited by bisindolylmaleimide I, a potent and specific inhibitor of PKC, but was unaffected by H89, an inhibitor of protein kinase A. In contrast, treatment with bisindolylmaleimide I did not affect the basal level of Nef phosphorylation, suggesting two different phosphorylation pathways. A PMA-insensitive CD4 mutant in which three serine residues in the cytoplasmic domain have been replaced by alanines was used to determine whether PMA-induced phosphorylation affects Nef-induced CD4 downregulation. In Nef-expressing cells, treatment with PMA enhanced downregulation of the CD4 serine triple mutant from the cell surface, suggesting that phosphorylation is important for Nef function.

CD4 Antigens↗
PubMed1997

[The significance of preoperative visual electrophysiology for vitrectomy].

OBJECTIVE: To investigate the prognostic value of preoperative visual electrophysiology in patients with vitreoretinopathy (VRP) who underwent vitrectomy. METHODS: Electroretinogram (ERG) and visual evoked potential (VEP) were performed prior to vitrectomy in 82 patients with vitreous hemorrhage or opacity due to ocular trauma, retinal detachment or retinal vascular diseases (Eales' disease, diabetic retinopathy, etc.). A comparative analysis was made of the effectiveness and visual acuities 2 weeks postoperatively. RESULTS: Among the 82 eyes, the preoperative ERG was normal in 6 eyes, slightly or moderately reduced in 37 eyes and severely reduced or had no response in 39 eyes. Of these 39 eyes, no improvement or reduction in postoperative visual acuity was seen in 23 and 6 eyes respectively, a sharp contrast to the postoperative results of the 6 normal ERG eyes. The preoperative ERGs differed due to different causes. A further analysis regarding the VEP of the patients who had poor postoperative results was made. CONCLUSIONS: The data showed that the preoperative visual electrophysiology was significantly correlated with the effectiveness of the operation. It is helpful in prognostic assessment of the postoperative outcome and visual acuity changes in VRP patients treated mainly by vitrectomy.

Electroretinography↗
PubMed1996

Inhibition of Nef- and phorbol ester-induced CD4 degradation by macrolide antibiotics.

Human immunodeficiency virus type 1 (HIV-1) is the causative agent of AIDS. The simian immunodeficiency virus (SIV) causes a similar syndrome in macaques. The product of the nef gene of SIV has been shown to be important for virus replication and disease progression in vivo. In vitro, both SIV and HIV Nef downregulate surface expression of CD4 and accelerate total CD4 turnover. The mechanism by which Nef downregulates CD4 has not been established. A current model suggests that Nef enhances cell surface CD4 endocytosis and degradation in lysosomes. However, this was recently challenged when CD4 was found to accumulate in early endosomes of cells expressing Nef. Because inhibition of Nef function might halt virus replication and disease progression, we tested two macrolide antibiotics for their ability to inhibit Nef function. Concanamycin B (ConB) and bafilomycin A1 (BFLA1) are specific inhibitors of acidification of cell endosomes and lysosomes and, unlike other inhibitors, do not affect transport. Although ConB (25 nM) and BFLA1 (100 nM) blocked phorbol myristate acetate- and Nef-induced CD4 degradation in human monocyte U937 cells, CD4 surface expression was not recovered. Instead, CD4 accumulated in lysosomes. To determine if Nef is directly responsible for CD4 degradation or if they bind to each other in a manner similar to Vpu, transcripts of human CD4 and HIV-1 nef were cotranslated in vitro. Our results indicate that under our experimental conditions, Nef does not affect CD4 stability and does not associate with CD4 in this in vitro system. Our data suggest that (i) CD4 downregulation by Nef results in degradation of CD4 in lysosomes, (ii) inhibition of CD4 degradation by macrolide antibiotics does not restore surface expression, and (iii) the inhibition of CD4 expression by Nef appears to be indirect and is likely to involve cellular factors.

Anti-Bacterial Agents↗
PubMed1996

The association of Nef with a cellular serine/threonine kinase and its enhancement of infectivity are viral isolate dependent.

The nef genes of human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) encode a 27- to 34-kDa myristoylated protein which induces downregulation of CD4 surface levels and enhances virus infectivity. In adult macaques, Nef has been implicated in pathogenesis and disease progression. Both HIV-1 SF2 Nef and SIVmac239 Nef have been shown to associate with a cellular serine/threonine kinase. We tested five functional Nef isolates to examine whether this kinase association is a property conserved among different isolates. HIV-1 SF2 and 248 and SIVmac239 Nef proteins were found associated with the kinase. HIV-1 NL4-3 and 233 Nef proteins were found weakly associated or not associated with the kinase. All five Nef isolates efficiently downregulated CD4 cell surface expression, suggesting that the association with this cellular kinase is not required for Nef to downregulate CD4. Comparison of the SF2 and NL4-3 isolates shows a differential ability of Nef to enhance infectivity that suggests a possible correlation between kinase association and enhancement of infectivity.

Animals↗
PubMed1994

A 55-kDa protein induced in Aedes albopictus (mosquito) cells by antiviral protein.

We have previously shown that the antiviral protein (AVP) produced by Sindbis virus-infected Aedes albopictus (mosquito) cells (Riedel and Brown, J. Virol. 29, 51-60, 1979) blocks Sindbis viral RNA synthesis and stimulates its own production when applied to uninfected A. albopictus cells (Luo and Brown, Virology 194, 44-49, 1993). From a virus-sensitive (wild-type) mosquito cell line (U4.4) we produced a virus-resistant cell line (L4.4) by exposing U4.4 cells to the AVP. L4.4 cells constitutively produce AVP and fail to replicate viral RNA after infection with virus or transfection with purified viral RNA. In this study we have compared cellular proteins produced as the sensitive cells are temporally converted to the resistant phenotype following exposure to the AVP. A 55-kDa membrane protein associated with lysosomes is found in L4.4 and not in U4.4 cells. This protein is induced during the first 48 hr following treatment of U4.4 cells with the AVP. The correlation of the appearance of the 55-kDa protein with the onset of virus resistance implies that this AVP-induced protein may be responsible for the inhibition of viral RNA replication. Although virus RNA synthesis is blocked in the L4.4 cell line, we have found that the synthesis of nonstructural proteins takes place.

Aedes↗
PubMed1993

Purification and characterization of a Sindbis virus-induced peptide which stimulates its own production and blocks virus RNA synthesis.

The antiviral activity described by Riedel and Brown (J. Virol. 29, 51-60) has been purified to homogeneity. A study of the effects of this pure peptide on mosquito cell growth and virus replication has been undertaken. We have found that treatment of noninfected Aedes albopictus cells with this antiviral peptide results in a temporary arrest of cell division accompanied by acquisition of virus resistance and the constitutive production of the antiviral protein.

Aedes↗
PubMed1992

The electrophysiology in idiopathic senile macular hole.

Of 12 patients with idiopathic senile full-thickness macular hole, 3 had bilateral involvement, 9 had monocular macular hole. Flash ERG and pattern VEP were performed in the bilateral eyes of all patients. The abnormal rate of the pattern VEP was 93.3% when we used 15' checkboard stimulus, the changes of the VEP appeared as delayed latencies, reduced amplitudes or malformation of P100. The abnormal rate of the flash ERG was 53.3%, showing primary characteristics of reduced amplitudes of cone response b-wave. There was statistically significant difference between the abnormal rates of the VEP and the ERG (P < 0.05). The halo diameters of the macular hole were significantly related to the latencies of P100 (P < 0.01), but there was no correlation between the hole diameter and the latencies of P100. In 9 fellow eyes which did not form macular hole, 3 eyes had the abnormal pattern VEP, including one eye having formed macular cyst. The results show that, in idiopathic senile macular hole, the pattern VEP provides more functional information than does the ERG, and prompts the importance of close follow-up of fellow eye.

Aged↗
PubMed1992

Quantitative evaluation of flicker ERG waveforms in low vision patients.

Flicker electroretinograms (FERGs) of 30Hz were recorded for 46 eyes (23 cases) of low vision, including 38 eyes with retinal diseases and 8 eyes with optic neuropathy, and for 39 normal eyes (33 cases). The recorded wave forms were analysed by two methods: (1) to measure the peak to peak amplitude and pseudophase directly, and (2) to measure the amplitude and phase of fundamental response component (30Hz) by discrete Fourier transform (DFT). The abnormality ratios between these two methods were compared in low vision patients. All the eyes with optic neuropathy showed normal flicker ERG, the abnormality ratios of directly measured amplitude and the amplitude by DFT were 60.9% in 28 eyes and 71.7% in 33 eyes with retinal diseases respectively, the difference being no statistically significant (P > 0.05), while the abnormality ratios of directly measured pseudo-phase and the phase by DFT were 60.9% in 28 eyes and 82.6% in 38 eyes with retinal diseases respectively, the difference being statistically significant (P < 0.05). These results suggest that FERGs are an objective way to test the retinal function, and DFT for flicker ERGs are more useful in quantitative evaluation of the retinal function.

Adolescent↗
PubMed1992

Visual functions and trace element metabolism in tobacco-toxic optic neuropathy.

Visual functions and nutrition metabolic characteristics were studied in 8 subjects (16 eyes) with tobacco-toxic optic neuropathy (TTON). Their visual functions tested by psychophysical and electrophysiologic methods showed that 1: 1. central vision diminished in 16 eyes, 2. dyschromatopsias were found in 14 tested eyes, 3. bilateral symmetrical central or cecocentral scotomas were the visual field characteristics in all cases, 4. PVEP were severe abnormal in 3 spatial frequencies in all cases and 56.3% of 15' checkboard PVEP showed flat responses, which indicated the impairment of optic nerve dominated by the central field. However, the preserved visual responses could be obtained by FVEP test in 14 tested eyes even though their visual acuity were between the range of 0.02-0.2 and flat PVEP responses. The II and III wave latencies of primary stage were more prolonged than those of control group (P < 0.01), which further indicated the preferential demyelination corresponding to the papillomacular bundles, 5. ERG showed slightly attenuated amplitudes in 5 of 8 tested eyes, which indicated the secondary and mild retinal lesion. On the other hand, TTON occurred on a background of long-term, heavy smoking, drinking, emaciation and malnutrition bodies with low serum zinc level.

Adult↗
PubMed1991

Abnormalities of ERG in congenital aniridia.

Congenital aniridia is generally associated with nystagmus, corneal pannus, cataract, ectopia lentis, glaucoma, macular hypoplasia, optic nerve hypoplasia and compromised visual function. Many theories have been proposed, including a failure in the development of the neural ectoderm and/or an aberrant development of mesoderm. We observed the ERG from 19 patients with congenital aniridia. Fourteen patients had abnormal ERG, including the reduced a wave trough under dark adapted red stimuli with dark adaptation; abnormal a or/and b waves in dark or/and light adapted ERG or abnormal oscillatory potentials. The ERG abnormalities reflected the disturbances either in the outer retina such as photoreceptors or in the structures of inner retina. The preserved visual function and several ocular abnormalities show that the aniridia may be a consequence of several pathogenetic mechanisms. This fact supported the opinion that congenital aniridia would be a concurrent failure of both ectoderm and mesoderm in embryonate development.

Adolescent↗
PubMed1991

The significance of testing preoperative visual function in cataract using laser interferometric visual acuity and ERG.

Tests of preoperative visual function and prediction of postoperative E chart visual acuity (ECVA) using laser interferometric visual acuity (LIVA) and electroretinogram (ERG) were performed in 16 cases (19 eyes) of cataract. The results showed that the coincident rate between preoperative LIVA and postoperative ECVA was 63.2%, and there was a parallel correlation between preoperative amplitude of photopic ERG b-wave and postoperative ECVA in 79.0% of the eyes. Comparing these two methods, the test of LIVA is very simple, fast, and is easily accepted by the patients with cataract. ERG is an objective method, which is slightly influenced by dense lens, but the test is more complex. Therefore, the combined use of these two methods will provide a more comprehensive and correct evaluation of preoperative visual function and a more reliable prediction for postoperative visual acuity in patients with cataract.

Adolescent↗
PubMed1991

[A clinical study and analysis of congenital lenticular dislocation (35 cases)].

Thirty-five cases of congenital lenticular dislocation seen in our Center since 1985 have been studied and analyzed clinically. By the survey of pedigrees and examination of these patients, including ocular, systemic, skeletal X-ray, psychocardiogram, and urinary sodium-nitroprusside test, 21 cases were diagnosed as Marfan's syndrome, 6 cases as simple ectopia lentis, 3 cases as Weill-Marchesani's syndrome, 4 cases as aniridia and 1 case as homecys tinuria. We found that the most significant ocular manifestation of congenital lenticular dislocation was reduction in visual acuity. The severity of visual disturbance varied with the types of dislocation and the visual deficiency was closely related to the intermediate-grade (II) dislocation of the lens. Examination of ERG showed normal function in most of the patients. From this, we believe that the major cause of visual reduction in congenital lenticular dislocation is lenticular myopia and astigmatism. There fore, early diagnosis and effective correction of vision should be emphasized to prevent the occurrence of amblyopia.

Amblyopia↗
PubMed1991

[Distribution and activity regularities of medicinal Mojiang centipedes in Yunnan Province].

A study has been made on the distribution, living environment and behavior characteristics of medicinal Mojiang centipedes in Yunnan Province. The result has shown they are active between mid-March and late October every year. During this time, the temperature is 18-23 degrees C, moisture 75-85 percent, precipitation 50-280mm, sunshine 115-225 hours and evaporation 100-200mm.

Animals↗