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Biomedical subjects

W Jiang

Publications and source records attributed to W Jiang.

At least 325 records · Page 18Linked to original sources
PubMed1993

Tissue-specific expression and chromosomal localization of the alpha subunit of mouse meprin A.

Meprins, membrane-bound oligomeric metalloendopeptidases, contain alpha and/or beta subunits. Their activities have been found in the mouse and rat kidney. The cloned cDNA for the mouse alpha subunit of meprin A (EC cloned cDNA for the mouse alpha subunit of meprin A (EC 3.4.24.18) was used here to survey mRNA expression in kidney of different mouse strains and in various tissues of mice and rats. A single message of 3.6 kilobases was found in kidney of random bred (ICR) and inbred mice (C57BL/6, DBA/2) that contain high meprin A activity and in Sprague-Dawley rat kidney. The alpha subunit message was undetectable in the kidney of C3H/He and CBA mice, inbred strains that do not express meprin A activity. Therefore, meprin A activity in the kidney of mouse strains correlates with the amount of alpha subunit mRNA present. The 3.6-kilobase mRNA meprin alpha subunit message was also detected in the small intestine of the rat but not in mice. No message was detected in brain, heart, skeletal muscle, liver, lung, or spleen of mice or rats. Polymerase chain reaction amplification or Southern blot analysis of genomic DNA revealed that the gene for the alpha subunit is present in all mouse strains as well as in human, monkey, rat, mouse, dog, cow, rabbit, and chicken, but it was not detected in yeast. There is one gene copy present in the mouse genome. The gene was localized to mouse chromosome 17 centromeric to the major histocompatibility complex (H-2) by the interspecific backcrossing method. The localization of this allele to Mep-1, the gene previously found to regulate the expression of meprin A activity in mice, supports the proposal that Mep-1 is the structural gene for the alpha subunit.

Animals↗
PubMed1993

Kinetics of binding and hydrolysis of a series of nucleoside triphosphates by actomyosin-S1. Relationship between solution rate constants and properties of muscle fibers.

We have measured the steady state kinetics of hydrolysis and presteady state kinetics of binding of the nucleoside triphosphate GTP, CTP, aza-ATP (1-N6-etheno-2-aza-ATP), and ATP by rabbit skeletal actomyosin-S1. The maximum rates of steady state hydrolysis at 10 degrees C at low ionic strength are: CTP, 1.9 s-1 > ATP, 1.3 s-1 > aza-ATP, 0.19 s-1 > GTP, 0.03 s-1. A similar dependence of the rate of steady state hydrolysis upon nucleotide structure has been observed in isometrically contracting muscle fibers in the accompanying paper (Pate, E., Franks-Skiba, K., White, H., and Cooke, R. (1993) J. Biol. Chem. 268, 10046-10053) which strongly suggests that the same biochemical step that limits the maximum rate of hydrolysis of nucleoside triphosphates by actomyosin-S1 in solution also limits the rate of hydrolysis by isometrically contracting muscle fibers. The apparent second order rate constants for the dissociation of actomyosin-S1 by nucleoside triphosphates at 10 degrees C are: ATP, 2.7 x 10(6) M-1 s-1 > aza-ATP, 3.4 x 10(5) M-1 s-1 > GTP, 2.5 x 10(5) M-1 s-1 > CTP, 1.4 x 10(5) M-1 s-1. There is an excellent correlation between the second order rate constant for the dissociation of actomyosin-S1 in solution and the dependence of shortening velocity in glycerinated muscle fibers upon the concentration for ATP, aza-ATP, and CTP (as per accompanying article; Pate et al., 1993). We have used the second order rate constants obtained in solution for the dissociation of actomyosin-S1 by these nucleotides and shortening velocity data obtained with the same nucleoside triphosphates in glycerinated psoas fibers in the accompanying article (Pate et al., 1993) to determine the average distance over which cross-bridges remain attached during unloaded shortening to be 5-12 nm.

Actins↗
PubMed1993

Correlation of p53 mutations with epidermal growth factor receptor overexpression and absence of mdm2 amplification in human esophageal carcinomas.

Esophageal carcinomas from 24 patients, most of whom were smokers and consumed alcoholic beverages daily, were analyzed for mutations in exons 5-8 of the p53 tumor suppressor gene. Mutations were identified by polymerase chain reaction amplification and direct sequencing in 12 of 24 (50%) of the samples; almost half of the mutations were at A:T base pairs. Nuclear accumulation of p53 protein, determined by immunohistochemistry with the CM-1 polyclonal antibody, was observed in all cases in which a missense mutation in the p53 gene was detected. None of the 24 carcinomas had amplification of the mdm2 gene, an alternate pathway to p53 loss of function. Alterations involving three other cancer-related genes associated with human esophageal carcinogenesis, c-erbB-1/epidermal growth factor receptor (EGFR), c-myc, and retinoblastoma (Rb), were examined by Southern blot or immunohistochemical analysis in the same sample set to explore the possibility of a link between oncogene activation and loss of tumor suppressor function. While no associations were observed between amplification of the c-myc or EGFR genes and p53 abnormalities, a significant correlation (P < 0.01) was seen between the presence of p53 mutation and EGFR overexpression. Absence of Rb protein, measured immunohistochemically, was observed in four tumors, none of which had aberrations of the p53 gene.

Carcinoma, Squamous Cell↗
PubMed1993

Effects of aluminium ion on root growth, cell division, and nucleoli of garlic (Allium sativum L.).

The effects of different concentrations of aluminium chloride on root growth, cell division, chromosome morphology and nucleoli in root tip cells of garlic (Allium sativum L.) were studied. The concentrations of aluminium chloride (AlCl(3)) used were 10(-5), 10(-4), 10(-3), 10(-2) and 10(-1) m. Aluminium chloride inhibited root growth and caused mitotic irregularities, including c-mitosis, anaphase bridges, and chromosome stickiness. Nucleolar material was extruded from the nucleus into the cytoplasm. Extrusion was observed in inner root meristem and root cap cells. The poisoning by Al(3+) of the root tip cells of Allium sativum may result from the uptake and accumulation of Al and inhibition of Ca uptake, distribution of physiological activities of calmodulin (CaM) and the inhibition of some enzyme reactions.

Journal Article↗
PubMed1993

Isolation and characterization of a gene (CBF2) specifying a protein component of the budding yeast kinetochore.

We have cloned and determined the nucleotide sequence of the gene (CBF2) specifying the large (110 kD) subunit of the 240-kD multisubunit yeast centromere binding factor CBF3, which binds selectively in vitro to yeast centromere DNA and contains a minus end-directed microtubule motor activity. The deduced amino acid sequence of CBF2p shows no sequence homologies with known molecular motors, although a consensus nucleotide binding site is present. The CBF2 gene is essential for viability of yeast and is identical to NDC10, in which a conditional mutation leads to a defect in chromosome segregation (Goh, P.-Y., and J. V. Kilmartin, in this issue of The Journal of Cell Biology). The combined in vitro and in vivo evidence indicate that CBF2p is a key component of the budding yeast kinetochore.

Amino Acid Sequence↗
PubMed1993

The backbone structure of the major cold-shock protein CS7.4 of Escherichia coli in solution includes extensive beta-sheet structure.

CS7.4 is the major cold-shock protein specifically expressed to a level as high as 13% of the total cellular protein within the first hour when Escherichia coli cell culture is shifted from 37 to 15 degrees C [Goldstein et al. (1990) Proc. Natl. Acad. Sci. USA 87, 283-287]. It consists of 70 amino acid residues with a very high content of aromatic residues. CS7.4 was overproduced and purified to homogeneity. Its secondary structure was analyzed by examining circular dichroism at both the far and near-UV regions; the results suggest that the protein is largely beta-sheet in conformation. The predominance of beta-sheet structure in the protein was confirmed by using Fourier-transform infrared spectroscopy. A folded compact conformation was also verified by fluorescence emission spectroscopy. We evaluated Tm, delta H, and delta S from the thermal denaturation profile of the protein. Unusual spectral features observed in the far-UV region are attributed to the high content of aromatic residues. The protein is relatively small and contains no disulfide bonds. However, it is surprisingly stable to heat denaturation.

Bacterial Proteins↗
PubMed1993

Chloramphenicol induces the transcription of the major cold shock gene of Escherichia coli, cspA.

A downshift in temperature or exposure of cells to certain inhibitors of translation has been shown to induce the synthesis of cold shock proteins in Escherichia coli. We characterized the induction of the major cold shock protein (CS7.4, the product of the cspA gene) of E. coli in response to a shift from 37 to 15 degrees C and in response to the addition of chloramphenicol at 15 degrees C. A pulse-labeling assay and primer extension experiments indicated that the cold shock treatment resulted in a transient increase in the level of the cspA transcript and a transient induction of CS7.4, while the addition of chloramphenicol resulted in a constitutive increase in the level of cspA transcript and constitutive production of CS7.4. The addition of rifamycin immediately following the temperature downshift or along with the addition of chloramphenicol repressed the transcription of cspA as well as the induced production of CS7.4. Furthermore, changes in the cspA mRNA level were coincident with changes in CS7.4 synthesis. These results indicate that the expression of cspA induced by cold shock and chloramphenicol is at the level of transcription but not at the level of translation. Measurement of the half-life revealed that the cspA mRNA induced by chloramphenicol was more stable than that induced by cold shock.

Bacterial Proteins↗
PubMed1993

An essential yeast protein, CBF5p, binds in vitro to centromeres and microtubules.

Yeast centromere DNA (CEN) affinity column chromatography has been used to purify several putative centromere and kinetochore proteins from yeast chromatin extracts. The single yeast gene (CBF5) specifying one of the major low-affinity centromere-binding proteins (p64'/CBF5p) has been cloned and shown to be essential for viability of Saccharomyces cerevisiae. CBF5 specifies a 55-kDa highly charged protein that contains a repeating KKD/E sequence domain near the C terminus, similar to known microtubule-binding domains in microtubule-associated proteins 1A and 1B, CBF5p, obtained by overexpression in bacterial cells, binds microtubules in vitro, whereas C-terminal deleted proteins lacking the (KKD/E)n domain do not. Dividing yeast cells containing a C-terminal truncated CBF5 gene, producing CBF5p containing only three copies of the KKD/E repeat, delay with replicated genomes at the G2/M phase of the cell cycle, while depletion of CBF5p arrests most cells in G1/S. Overproduction of CBF5p in S. cerevisiae complements a temperature sensitivity mutation in the gene (CBF2) specifying the 110-kDa subunit of the high-affinity CEN DNA-binding factor CBF3, suggesting in vivo interaction of CBF5p and CBF3. A second low-affinity centromere-binding factor has been identified as topoisomerase II.

Base Sequence↗
PubMed1993

Gaze-related activity of putative inhibitory burst neurons in the head-free cat.

1. Previous studies in the cat have demonstrated that output neurons of the superior collicular as well as brain stem omnipause neurons have discharges that are best correlated, not with the trajectory of the eye in the head but, with the trajectory of the visual axis in space (gaze = eye-in-head + head-in-space) during rapid orienting coordinated eye and head movements. In this study, we describe the gaze-related activity of cat premotor "inhibitory burst neurons" (IBNs) identified on the basis of their position relative to the abducens nucleus. 2. The firing behavior of IBNs was studied during 1) saccades made with the head stationary, 2) active orienting combined eye-head gaze shifts, and 3) passive movements of the head on the body. IBN discharges were well correlated with the duration and amplitude of saccades made when the head was stationary. In both head-free paradigms, the behavior of cat IBNs differed from that of previously described primate "saccade bursters". The duration of their burst was better correlated with gaze than saccade duration, and the total number of spikes in a burst was well correlated with gaze amplitude and generally poorly correlated with saccade amplitude. The behavior of cat IBNs also differed from that of previously described primate "gaze bursters". The slope of the relationship between the total number of spikes and gaze amplitude observed during head-free gaze shifts was significantly lower than that observed during head-fixed saccades. 3. These studies suggest that cat IBNs do not fit into the categories of gaze-bursters or saccade-bursters that have been described in primate studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Abducens Nerve↗
PubMed1993

[The genotyping of HLA-D region and specific RFLP patterns for Chinese Han population].

The HLA class II antigens play very an important role in presenting foreign antigens. The class II gene typing at the DNA level possesses higher capability to recognize polymorphism so that it can disclose new specificities. According to the RFLP patterns of DRB and DQA genes for Caucasian HTC and some Chinese homozygous cells, we typed 90 randomly sampled healthy Chinese Hans in Shanghai area, and compared the results obtained with those from serological typing. In addition to being reliable and superior to serological typing, we found some unique RFLP patterns of the HLA-D region in Chinese Han population based on comparison with Caucasian patterns.

Blood Grouping and Crossmatching↗
PubMed1993

Overexpression of cyclin D1 in rat fibroblasts causes abnormalities in growth control, cell cycle progression and gene expression.

Cyclin D1, a putative G1 cyclin, has been implicated in cell cycle control. The human cyclin D1 gene is located on chromosome 11q13 where DNA rearrangement and amplification have been detected in several types of human cancer. Previous studies demonstrated that the cyclin D1 gene is not only rearranged or amplified but also overexpressed in some of these human tumors and tumor-derived cell lines. To further address the roles of cyclin D1 in cell cycle control and tumorigenesis, we have stably overexpressed the human cyclin D1 cDNA in Rat6 embryo fibroblasts by using retrovirus mediated transduction. The cyclin D1 protein was overproduced about 10-fold and was localized predominately in the nucleus. Cyclin D1 overexpressing cells displayed a decrease in the duration of the G1 phase, decreased cell size, and induced tumors when injected into athymic (nude) mice. In addition, overexpression of cyclin D1 in Rat6 cells perturbed the expression of several cellular growth-related genes including c-myc, c-jun, and cyclin A, but not cyclin D3. Taken together, these results indicate that deregulated expression of the cyclin D1 gene can cause disturbances in cell cycle control and gene expression and also enhance tumorigenesis.

Animals↗