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H Fu

Publications and source records attributed to H Fu.

At least 127 records · Page 7Linked to original sources
PubMed1996

The multiubiquitin-chain-binding protein Mcb1 is a component of the 26S proteasome in Saccharomyces cerevisiae and plays a nonessential, substrate-specific role in protein turnover.

The 26S proteasome is an essential proteolytic complex that is responsible for degrading proteins conjugated with ubiquitin. It has been proposed that the recognition of substrates by the 26S proteasome is mediated by a multiubiquitin-chain-binding protein that has previously been characterized in both plants and animals. In this study, we identified a Saccharomyces cerevisiae homolog of this protein, designated Mcb1. Mcb1 copurified with the 26S proteasome in both conventional and nickel chelate chromatography. In addition, a significant fraction of Mcb1 in cell extracts was present in a low-molecular-mass form free of the 26S complex. Recombinant Mcb1 protein bound multiubiquitin chains in vitro and, like its plant and animal counterparts, exhibited a binding preference for longer chains. Surprisingly, (delta)mcb1 deletion mutants were viable, grew at near-wild-type rates, degraded the bulk of short-lived proteins normally, and were not sensitive to UV radiation or heat stress. These data indicate that Mcb1 is not an essential component of the ubiquitin-proteasome pathway in S.cerevisiae. However, the (delta)mcb1 mutant exhibited a modest sensitivity to amino acid analogs and had increased steady-state levels of ubiquitin-protein conjugates. Whereas the N-end rule substrate, Arg-beta-galactosidase, was degraded at the wild-type rate in the (delta)mcb1 strain, the ubiquitin fusion degradation pathway substrate, ubiquitin-Pro-beta-galactosidase, was markedly stabilized. Collectively, these data suggest that Mcb1 is not the sole factor involved in ubiquitin recognition by the 26S proteasome and that Mcb1 may interact with only a subset of ubiquitinated substrates.

Amino Acid Sequence↗
PubMed1996

Cancer mortality among shoe manufacturing workers: an analysis of two cohorts.

OBJECTIVES: To examine the cancer risk of shoe manufacturing workers and evaluate whether the risk was associated with exposure to leather dust and solvents. METHODS: Data from two historical cohort studies of shoe workers were expanded and analysed in parallel. A total of 4215 shoemakers from England contributing 103 726 person-years at risk and 2008 shoemakers from Florence, Italy, contributing 54,395 person-years at risk were included in the analysis. Exposure to leather dusts and solvents from glues was evaluated on the basis of job title information. Standardised mortality ratios (SMR) were calculated as ratios of observed deaths (Obs) over expected derived from national mortalities. RESULTS: Overall mortality was lower than expected in both cohorts (English cohort: Obs 3314, SMR 81, 95% confidence interval (95% CI) 78-84; Florence cohort: Obs 333, SMR 87, 95% CI 78-97). An increased risk of nasal cancer was found (English cohort: Obs 12, SMR 741; Florence cohort: Obs 1, SMR 909). 10 of the 13 cases occurred among English workers employed in the manufacture of welted boots (SMR 926, 95% CI 444-1703), a sector of the industry thought to have had the highest exposure to leather dust. Mortality from leukaemia was not increased in the English cohort (Obs 16, SMR 89), but was increased in the Florence cohort (Obs 8, SMR 214, 95% CI 92-421); and the highest risk was found among shoe workers in Florence who were first exposed between 1950 and 1959 when exposure to benzene was substantial (Obs 3, SMR 536, 95% CI 111-1566). Some evidence for an excess risk of stomach, bladder, and kidney cancer, as well as multiple myeloma and non-Hodgkin's lymphoma was also found in the Florence cohort only among workers employed in jobs with the highest exposure to solvents. CONCLUSIONS: These findings confirm the associations between exposure to leather dust and nasal cancer and between exposure to benzene and leukaemia in the shoe manufacturing industry and suggest that the risk of other cancers may be increased among workers exposed to solvents or glues.

Adult↗
PubMed1995

Crystal structure of the zeta isoform of the 14-3-3 protein.

The 14-3-3 family of proteins have recently been identified as regulatory elements in intracellular signalling pathways: 14-3-3 proteins bind to oncogene and proto-oncogene products, including c-Raf-1 (refs 2-5), c-Bcr (ref. 6) and polyomavirus middle-T antigen; overexpression of 14-3-3 activates Raf kinase in yeast and induces meiotic maturation in Xenopus oocytes. Here we report the crystal structure of the major isoform of mammalian 14-3-3 proteins at 2.9 A resolution. Each subunit of the dimeric protein consists of a bundle of nine antiparallel helices that form a palisade around an amphipathic groove. The groove is large enough to accommodate a tenth helix, and we propose that binding to an amphipathic helix represents a general mechanism for the interaction of 14-3-3 with diverse cellular proteins. The residues in the dimer interface and the putative ligand-binding surface are invariant among vertebrates, yeast and plants, suggesting a conservation of structure and function throughout the 14-3-3 family.

14-3-3 Proteins↗
PubMed1995

Sink- and vascular-associated sucrose synthase functions are encoded by different gene classes in potato.

Two differentially regulated classes of sucrose synthase genes, Sus3 and Sus4, were identified in potato. They cannot be classified as Sus1 and Sus2 types based on sequence homology and appear to have evolved after the divergence of the major families of dicotyledonous plants but before the divergence of tomato and potato. The potato sucrose synthase clones Sus3-65 and Sus4-16 share an 87% nucleotide identity in the coding regions, and both are interrupted by 13 introns, including a long leader intron. Potato Sus3 genes are expressed at the highest levels in stems and roots and appear to provide the vascular function of sucrose synthase. In contrast, Sus4 genes are expressed primarily in the storage and vascular tissue of tubers and appear to facilitate sink function. The genes are differentially regulated in root tips, with Sus3 expressed at high levels in the cell division zone and Sus4 expressed at high levels in the meristem and cap.

Amino Acid Sequence↗
PubMed1995

High-level tuber expression and sucrose inducibility of a potato Sus4 sucrose synthase gene require 5' and 3' flanking sequences and the leader intron.

The 3.6 kb of 5' flanking sequence, leader intron, and 0.7 kb of 3' sequence from the potato sucrose synthase gene Sus4-16 are sufficient to direct high-level expression in developing tubers, in basal tissues of axillary buds and shoots, and in meristems and caps of roots, and to confer sucrose inducibility in leaves. By examining a series of deletion and substitution constructs in transgenic potato plants, we found that this pattern of expression requires 5' flanking sequences both upstream and downstream of position -1500 and that sequences between positions -1500 and -267 are essential for sucrose induction. Replacement of the native 3' sequence with the nopaline synthase 3' sequence resulted in the loss of sucrose inducibility and of expression in basal tissues of axillary buds. A general decrease in expression in other tissues was also observed. Removal of the 1612-bp leader intron also had a dramatic effect on both the pattern and level of expression.

Base Sequence↗
PubMed1995

A potato Sus3 sucrose synthase gene contains a context-dependent 3' element and a leader intron with both positive and negative tissue-specific effects.

To examine which sequences are involved in regulating the potato sucrose synthase gene Sus3-65, we examined a series of deletion and substitution constructs in transgenic potato and tobacco plants. In a construct containing 3.9 kb of 5' flanking region, substitution of the native 3' sequence with the nopaline synthase 3' sequence and deletion of the leader intron did not significantly affect expression in vegetative tissues. However, in a construct containing only 320 bp of 5' flanking region, these changes had marked effects. Replacing the native 3' sequences with nopaline synthase 3' sequences caused a six- to 20-fold increase in expression in vascular tissue, and removing the leader intron almost completely abolished expression in potato plants. Surprisingly, removal of the leader intron from either the full-length construct or a construct containing only 320 bp of 5' flanking sequence reduced expression in vascular tissue of tobacco anthers at later stages of development but increased expression in pollen by more than 100-fold.

Base Sequence↗
PubMed1995

Cancer and occupational exposure to inorganic lead compounds: a meta-analysis of published data.

OBJECTIVES: To review and summarise the epidemiological evidence on the carcinogenicity of occupational exposure to inorganic lead. METHODS: Case-control and cohort studies were reviewed and combined for meta-analysis. Fixed and random effect methods were used to estimate the summary effects. RESULTS: The combined results show a significant excess risk of overall cancer, stomach cancer, lung cancer, and bladder cancer, with relative risk ratios (RRs) and 95% confidence intervals (95% CIs) in the meta-analysis of 1.11 (1.05-1.17), 1.33 (1.18-1.49), 1.29 (1.10-1.50), and 1.41 (1.16-1.71) respectively. The RR (95% CI) for kidney cancer was also high, but did not reach significance (1.19 (0.96-1.48)). A separate analysis of studies of heavily exposed workers provided slightly increased RRs for cancers of the stomach (1.50) and lung (1.42). CONCLUSIONS: The findings from the workers with heavy exposure to lead provided some evidence to support the hypothesis of an association between stomach and lung cancer and exposure to lead. The main limitation of the present analysis is that the excess risks do not take account of potential confounders, because little information was available for other occupational exposures, smoking, and dietary habits. To some extent, the risk of lung cancer might be explained by confounders such as tobacco smoking and exposure to other occupational carcinogens. The excess risk of stomach cancer may also be explained, at least in part, by non-occupational factors. For bladder and kidney cancers, the excess risks are only suggestive of a true effect because of possible publication bias.

Case-Control Studies↗
PubMed1995

[Survival rates in patients with systemic lupus erythematosus].

To assess the impact of demographic and clinical factors on prognosis in patients with systemic lupus erythematosus (SLE), we examined the survival rate by lifetable analysis in 566 patients. All patients were Shanghai citizens who were diagnosed as having SLE in Huashan Hospital between 1959 and 1992. According to American Rheumatism Association's preliminary criteria. The survival rate from the time of SLE onset was 93% at 1 year, 73% at 5 years and 60% at 10 years. On univariate analysis, we found that the following factors worsened the probability of survival; male, neuropsychiatric manifestation, pleurisy-pericarditis, anemia, thrombocytopenia, lymphocytopenia, proteinuria, hematuria, urinary cast, azotemia, decreased endogenous creatinine clearance, increased cholesterol in serum, hypocomplementemia, abnormal electrocardiograph and high corticosteroid dose of treatment. On multivariate analysis, we found the four independent risk factors were male, azotemia, hypocomplementemia and high corticosteroid dose of treatment.

Adolescent↗
PubMed1994

Engineered biosynthesis of novel polyketides: influence of a downstream enzyme on the catalytic specificity of a minimal aromatic polyketide synthase.

To identify the minimum set of polyketide synthase (PKS) components required for in vivo biosynthesis of aromatic polyketides, combinations of genes encoding subunits of three different aromatic PKSs--act from Streptomyces coelicolor A3(2) (an actinorhodin producer), fren from Streptomyces roseofulvus (a frenolicin and nanaomycin producer), and tcm from Streptomyces glaucescens (a tetracenomycin producer)--were expressed in a recently developed Streptomyces host-vector system. The "minimal" components (ketosynthase/putative acyltransferase, chain length-determining factor, and acyl carrier protein) were produced with and without a functional polyketide ketoreductase and/or cyclase, and the polyketide products of these recombinant strains were structurally characterized. Several previously identified polyketides were isolated in addition to two previously unidentified polyketides, dehydromutactin and SEK 15b, described here. The results proved that the act cyclase is not required for the biosynthesis of several aberrantly cyclized products that have been previously reported. They are also consistent with earlier conclusions that the minimal PKS controls chain length as well as the regiospecificity of the first cyclization and that it can do so in the absence of both a ketoreductase and a cyclase. However, the ability of the minimal tcm PKS to synthesize two different singly cyclized intermediates suggests that it is unable to accurately control the course of this reaction by itself. In the presence of a downstream enzyme, the flux through one branch of the cyclization pathway increases relative to the other. We propose that these alternative specificities may be due to the ability of downstream enzymes to associate with the minimal PKS and to selectively inhibit a particular branch of the cyclization pathway.

DNA Mutational Analysis↗
PubMed1994

Interaction of the protein kinase Raf-1 with 14-3-3 proteins.

Members of a family of highly conserved proteins, termed 14-3-3 proteins, were found by several experimental approaches to associate with Raf-1, a central component of a key signal transduction pathway. Optimal complex formation required the amino-terminal regulatory domain of Raf-1. The association of 14-3-3 proteins and Raf-1 was not substantially affected by the activation state of Raf.

14-3-3 Proteins↗
PubMed1994

Association of the protein kinases c-Bcr and Bcr-Abl with proteins of the 14-3-3 family.

In this study, a protein that interacts with sequences encoded by the first exon of the protein kinase Bcr was cloned. The Bcr-associated protein 1 (Bap-1) is a member of the 14-3-3 family of proteins. Bap-1 interacts with full-length c-Bcr and with the chimeric Bcr-Abl tyrosine kinase of Philadelphia chromosome (Ph1)-positive human leukemias. Bap-1 is a substrate for the Bcr serine-threonine kinase and is also phosphorylated on tyrosine by Bcr-Abl but not by c-Abl. Bap-1 may function in the regulation of c-Bcr and may contribute to the transforming activity of Bcr-Abl in vivo. 14-3-3 proteins are essential for cell proliferation and have a role in determining the timing of mitosis in yeast. Through direct binding to sequences present in Bcr and in other proteins implicated in signaling, the mammalian 14-3-3 proteins may link specific signaling protein components to mitogenic and cell-cycle control pathways.

14-3-3 Proteins↗
PubMed1994

Waiting for radiotherapy in Ontario.

PURPOSE: Waiting lists for radiotherapy are a fact of life at many Canadian cancer centers. The purpose of this study was to provide a detailed description of the magnitude of the problem in Ontario. METHODS AND MATERIALS: The interval between diagnosis and initiation of radiation treatment was calculated for all patients receiving primary radiotherapy for carcinoma of the larynx, cervix, lung, and prostate at seven Ontario cancer centers between 1982 and 1991. The interval between surgery and initiation of postoperative radiotherapy for breast cancer was also calculated over the same period. The intervals between diagnosis and referral (t1), between referral and consultation (t2), and between consultation and initiation of radiotherapy (t3), were analyzed separately to determine where delay occurred. RESULTS: Median waiting times between diagnosis and initiation of radical treatment for carcinoma of the larynx, carcinoma of the cervix, nonsmall cell lung cancer, and carcinoma of the prostate were 30.3 days, 27.2 days, 27.3 days, and 93.3 days, respectively. The exceptional interval between diagnosis and treatment of prostate cancer was due to much longer delays between diagnosis and referral. The median waiting time between diagnosis and initiation of postoperative radiotherapy for breast cancer was 61.4 days and the median time between the completion of surgery and initiation of postoperative radiotherapy was 57.8 days. There were significant intercenter variations in median waiting times, but in every situation the median waiting time in Ontario as a whole increased steadily between 1982 and 1991. Median waiting times from diagnosis to the start of curative treatment for laryngeal cancer, cervical cancer, nonsmall cell lung cancer, and prostate cancer increased by 178.7%, 105.6%, 158.3%, and 62.9%, respectively. Waiting time from completion of surgery to initiation of postoperative radiotherapy for breast cancer increased by 102.7%. Most of the increase in treatment delay was found in the interval between consultation and initiation of radiotherapy. CONCLUSIONS: The Committee on Standards of the Canadian Association of Radiation Oncologists recommends that the interval between referral and consultation should not exceed 2 weeks and that the interval between consultation and initiation of radiotherapy should also not exceed 2 weeks. The majority of patients treated in Ontario met both those standards in 1982, but by 1991 few patients received care within the prescribed intervals.

Breast Neoplasms↗
PubMed1994

Engineered biosynthesis of novel polyketides: stereochemical course of two reactions catalyzed by a polyketide synthase.

A genetically engineered strain expressing the essential components of the tetracenomycin polyketide synthase (tcm PKS) along with the actinorhodin ketoreductase (act KR) was found to produce two new (diastereomeric) aromatic polyketides, designated RM20b and RM20c, in addition to RM20, whose structure was reported earlier [McDaniel, R., Ebert-Khosla, S., Hopwood, D. A., & Khosla, C. (1993) Science 262, 1546-1550]. Spectroscopic and in vivo isotopic labeling analysis of RM20b and RM20c revealed that their polyketide backbones were identical to that of RM20 with respect to chain length, regiospecificity of ketoreduction, and regiospecificity of the first intramolecular aldol condensation. This is consistent with earlier predictions that the essential components of the PKS--a bifunctional ketosynthase/acyltransferase, a chain length determining factor, and an acyl carrier protein--are responsible for controlling each of these features of the polyketide backbone [McDaniel, R., Ebert-Khosla, S., Hopwood, D. A., & Khosla, C. (1993) Science 262, 1546-1550; McDaniel, R., Ebert-Khosla, S., Hopwood, D. A., & Khosla, C. (1993) J. Am. Chem. Soc. 115, 11671-11675; Fu, H., Ebert-Khosla, S., Hopwood, D. A., & Khosla, C. (1994) J. Am. Chem. Soc. 116, 4166-4170]. In addition, however, RM20b and RM20c possess two unusual features. In both molecules the hydroxyls on sp3 C-9 and C-7 of the first six-membered ring, which arise as a result of ketoreduction and aldol condensation, respectively, are intact, rather than being lost via dehydration. Furthermore, the relative yield of RM20b (in which these hydroxyls are syn) is 7-fold greater than that of RM20c (in which they are anti).(ABSTRACT TRUNCATED AT 250 WORDS)

Carbon Isotopes↗
PubMed1994

Association of polyomavirus middle tumor antigen with 14-3-3 proteins.

To carry out its transformation function, the middle tumor antigen (MT) of murine polyomavirus associates with a number of cellular proteins involved in regulation of cell proliferation, including pp60c-Src, phosphatidylinositol 3-kinase, protein phosphatase 2A, Src homologous and collagen protein and growth factor receptor-binding protein 2. Here, two additional MT-associated proteins were identified as members of the 14-3-3 family of proteins. Yeast homologs of 14-3-3 proteins have recently been shown to play a role in the timing of mitosis. Thus, regulation of 14-3-3 protein function by MT may contribute to the development of neoplasia.

14-3-3 Proteins↗
PubMed1994

Lung cancer among tin miners in southeast China: silica exposure, silicosis, and cigarette smoking.

The Dachang Tin Mine is an industrial facility with high rates of lung cancer compared to the resident population in Guangxi province in southern China. Historically, exposure to silica dust was very high in the 1950s, falling in recent years to levels in keeping with international standards. Radon levels in the mine are low. We report the findings of a case-control study nested into a cohort study on miners working in Dachang. Cases of lung cancer among miners incident from 1973-1989 were obtained from local comprehensive medical records covering workers employed at the mine. These were matched approximately 3 to 1 with miners randomly chosen from the district surrounding the mine within the same birth decade. Matched odds ratios of 2.42 (95% confidence limit [CL] 1.3, 4.4) for underground employment, 3.52 (95% CL 1.7, 7.5) for smoking, and 2.04 (95%) CL 1.2, 3.7) for silicosis as determined on chest film were noted. Multifactor analysis of unconditional logistic regression showed that among the risk factors for excess mortality from lung cancer only the years spent drilling underground and the cumulative smoking index (product of daily cigarette consumption and number of years smoking) were independent contributors to risk and there was no interaction observed. The presence of silicosis did not contribute to predicting risk independently of the years spent underground.

Case-Control Studies↗
PubMed1994

A single Arabidopsis GF14 isoform possesses biochemical characteristics of diverse 14-3-3 homologues.

Arabidopsis cDNA clones of GF14 proteins originally were isolated on the basis of their association with the G-box DNA/protein complex by a monoclonal antibody screening approach. GF14 proteins are homologous to the 14-3-3 family of mammalian proteins. Here we demonstrate that recombinant GF14 omega, one member of the Arabidopsis GF14 protein family, is a dimeric protein that possesses many of the attributes of diverse mammalian 14-3-3 homologues. GF14 omega activates rat brain tryptophan hydroxylase and protein kinase C in a manner similar to the bovine 14-3-3 protein. It also activates exoenzyme S of Pseudomonas aeruginosa as does bovine brain factor activating exoenzyme S (FAS), which is itself a member of 14-3-3 proteins. In addition, GF14 omega binds calcium, as does the human 14-3-3 homologue reported to be a phospholipase A2. These results indicate that a single isoform of this plant protein family can have multiple functions and that individual GF14 isoforms may have multiple roles in mediating signal transductions in plants. However, GF14 omega does not regulate growth in an in vivo test for functional similarity to the yeast 14-3-3 homologue, BMH1. Thus, while a single plant GF14 isoform can exhibit many of the biochemical attributes of diverse mammalian 14-3-3 homologues, open questions remain regarding the physiological functions of GF14/14-3-3 proteins.

14-3-3 Proteins↗