排序方式: 共有56条查询结果,搜索用时 31 毫秒
1.
A. de Groot J. -J. Krijger A. Filloux J. Tommassen 《Molecular & general genetics : MGG》1996,250(4):491-504
InPseudomonas aeruginosa, the products of thexcp genes are required for the secretion of exoproteins across the outer membrane. Despite structural conservation of the Xcp components, secretion of exoproteins via the Xcp pathway is generally not found in heterologous organisms. To study the specificity of this protein secretion pathway, thexcp genes of another fluorescent pseudomonad, the plant growth-promotingPseudomonas putida strain WCS358, were cloned and characterized. Nucleotide sequence analysis revealed the presence of at least five genes, i.e.,xcpP, Q, R, S, andT, with homology toxcp genes ofP. aeruginosa. Unlike the genetic organization inP. aeruginosa, where thexcp cluster consists of two divergently transcribed operons, thexcp genes inP. putida are all oriented in the same direction, and probably comprise a single operon. Upstream ofxcpP inP. putida, an additional open reading frame, with no homolog inP. aeruginosa, was identified, which possibly encodes a lipoprotein. Mutational inactivation ofxcp genes inP. putida did not affect secretion, indicating that no proteins are secreted via the Xcp system under the growth conditions tested, and that an alternative secretion system is operative. To obtain some insight into the secretory pathway involved, the amino acid sequence of the N-terminus of the major extracellular protein was determined. The protein could be identified as flagellin. Mutations in thexcpQ andR genes ofP. aeruginosa could not be complemented by introduction of the correspondingxcp genes ofP. putida. However, expression of a hybrid XcpR protein, composed of the N-terminal one-third ofP. aeruginosa XcpR and the C-terminal two-thirds ofP. putida XcpR, did restore protein secretion in aP. aeruginosa xcpR mutant. 相似文献
2.
G.C. Krijger H.A. Claessens H. Th. Wolterbeek 《Chemical Speciation and Bioavailability》2013,25(1-2):29-36
ABSTRACTThe present paper addresses the potential use of capillary zone electrophoresis (CZE) as an analytical tool in 99- technetium speciation studies. In order to optimise sampling, storage and analytical procedures, the three marker compounds 99mTcO4-, 99mTc-DTPA and 99mTc-citrate were synthesised and used in test-measurements with CZE. The results underline the superior separation power of the CZE technique, and indicate good CZE performance for the stable 99mTcO4- and 99mTc-DTPA compounds. The data suggest that CZE may be used without problems for various Tc-compounds of intermediate mobilities. The specific data of 99mTc-citrate suggest that with this marker compound a threshold lability is reached for the use of CZE in plant Tc-speciation studies. This result means that CZE cannot be used in analyses of Tc-compounds which are less stable than Tc-citrate. Future CZE work will comprise the synthesis and use of Tc-markers of intermediate mobilities and stabilities; furthermore, effects of marker matrices and the plant matrix on CZE performance will be investigated. 相似文献
3.
Transcriptome analysis of root transporters reveals participation of multiple gene families in the response to cation stress 总被引:31,自引:4,他引:27
4.
Regulation of primary carbon metabolism in Kluyveromyces lactis 总被引:2,自引:0,他引:2
Breunig KD Bolotin-Fukuhara M Bianchi MM Bourgarel D Falcone C Ferrero I Frontali L Goffrini P Krijger JJ Mazzoni C Milkowski C Steensma HY Wésolowski-Louvel M Zeeman AM 《Enzyme and microbial technology》2000,26(9-10):771-780
In the recent past, through advances in development of genetic tools, the budding yeast Kluyveromyces lactis has become a model system for studies on molecular physiology of so-called “Nonconventional Yeasts.” The regulation of primary carbon metabolism in K. lactis differs markedly from Saccharomyces cerevisiae and reflects the dominance of respiration over fermentation typical for the majority of yeasts. The absence of aerobic ethanol formation in this class of yeasts represents a major advantage for the “cell factory” concept and large-scale production of heterologous proteins in K. lactis cells is being applied successfully. First insight into the molecular basis for the different regulatory strategies is beginning to emerge from comparative studies on S. cerevisiae and K. lactis. The absence of glucose repression of respiration, a high capacity of respiratory enzymes and a tight regulation of glucose uptake in K. lactis are key factors determining physiological differences to S. cerevisiae. A striking discrepancy exists between the conservation of regulatory factors and the lack of evidence for their functional significance in K. lactis. On the other hand, structurally conserved factors were identified in K. lactis in a new regulatory context. It seems that different physiological responses result from modified interactions of similar molecular modules. 相似文献
5.
Lindsey Oudijk Francien van Nederveen Cécile Badoual Frédérique Tissier Arthur S. Tischler Marcel Smid José Gaal Charlotte Lepoutre-Lussey Anne-Paule Gimenez-Roqueplo Winand N.M. Dinjens Esther Korpershoek Ronald de Krijger Judith Favier 《PloS one》2015,10(3)
Pheochromocytomas (PCCs) are neuroendocrine tumors arising from chromaffin cells of the adrenal medulla. Related tumors that arise from the paraganglia outside the adrenal medulla are called paragangliomas (PGLs). PCC/PGLs are usually benign, but approximately 17% of these tumors are malignant, as defined by the development of metastases. Currently, there are no generally accepted markers for identifying a primary PCC or PGL as malignant. In 2002, Favier et al. described the use of vascular architecture for the distinction between benign and malignant primary PCC/PGLs. The aim of this study was to validate the use of vascular pattern analysis as a test for malignancy in a large series of primary PCC/PGLs. Six independent observers scored a series of 184 genetically well-characterized PCCs and PGLs for the CD34 immunolabeled vascular pattern and these findings were correlated to the clinical outcome. Tumors were scored as malignant if an irregular vascular pattern was observed, including vascular arcs, parallels and networks, while tumors with a regular pattern of short straight capillaries were scored as benign. Mean sensitivity and specificity of vascular architecture, as a predictor of malignancy was 59.7% and 72.9%, respectively. There was significant agreement between the 6 observers (mean κ = 0.796). Mean sensitivity of vascular pattern analysis was higher in tumors >5 cm (63.2%) and in genotype cluster 2 tumors (100%). In conclusion, vascular pattern analysis cannot be used in a stand-alone manner as a prognostic tool for the distinction between benign and malignant PCC, but could be used as an indicator of malignancy and might be a useful tool in combination with other morphological characteristics. 相似文献
6.
PCNA ubiquitination is important, but not essential for translesion DNA synthesis in mammalian cells
Hendel A Krijger PH Diamant N Goren Z Langerak P Kim J Reissner T Lee KY Geacintov NE Carell T Myung K Tateishi S D'Andrea A Jacobs H Livneh Z 《PLoS genetics》2011,7(9):e1002262
Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism in which specialized low-fidelity DNA polymerases bypass replication-blocking lesions, and it is usually associated with mutagenesis. In Saccharomyces cerevisiae a key event in TLS is the monoubiquitination of PCNA, which enables recruitment of the specialized polymerases to the damaged site through their ubiquitin-binding domain. In mammals, however, there is a debate on the requirement for ubiquitinated PCNA (PCNA-Ub) in TLS. We show that UV-induced Rpa foci, indicative of single-stranded DNA (ssDNA) regions caused by UV, accumulate faster and disappear more slowly in Pcna(K164R/K164R) cells, which are resistant to PCNA ubiquitination, compared to Pcna(+/+) cells, consistent with a TLS defect. Direct analysis of TLS in these cells, using gapped plasmids with site-specific lesions, showed that TLS is strongly reduced across UV lesions and the cisplatin-induced intrastrand GG crosslink. A similar effect was obtained in cells lacking Rad18, the E3 ubiquitin ligase which monoubiquitinates PCNA. Consistently, cells lacking Usp1, the enzyme that de-ubiquitinates PCNA exhibited increased TLS across a UV lesion and the cisplatin adduct. In contrast, cells lacking the Rad5-homologs Shprh and Hltf, which polyubiquitinate PCNA, exhibited normal TLS. Knocking down the expression of the TLS genes Rev3L, PolH, or Rev1 in Pcna(K164R/K164R) mouse embryo fibroblasts caused each an increased sensitivity to UV radiation, indicating the existence of TLS pathways that are independent of PCNA-Ub. Taken together these results indicate that PCNA-Ub is required for maximal TLS. However, TLS polymerases can be recruited to damaged DNA also in the absence of PCNA-Ub, and perform TLS, albeit at a significantly lower efficiency and altered mutagenic specificity. 相似文献
7.
Monoubiquitylation of the homotrimeric DNA sliding clamp PCNA at lysine residue 164 (PCNA(K164)) is a highly conserved, DNA damage-inducible process that is mediated by the E2/E3 complex Rad6/Rad18. This ubiquitylation event recruits translesion synthesis (TLS) polymerases capable of replicating across damaged DNA templates. Besides PCNA, the Rad6/Rad18 complex was recently shown in yeast to ubiquitylate also 9-1-1, a heterotrimeric DNA sliding clamp composed of Rad9, Rad1, and Hus1 in a DNA damage-inducible manner. Based on the highly similar crystal structures of PCNA and 9-1-1, K185 of Rad1 (Rad1(K185)) was identified as the only topological equivalent of PCNA(K164). To investigate a potential role of posttranslational modifications of Rad1(K185) in DNA damage management, we here generated a mouse model with a conditional deletable Rad1(K185R) allele. The Rad1(K185) residue was found to be dispensable for Chk1 activation, DNA damage survival, and class switch recombination of immunoglobulin genes as well as recruitment of TLS polymerases during somatic hypermutation of immunoglobulin genes. Our data indicate that Rad1(K185) is not a functional counterpart of PCNA(K164). 相似文献
8.
Parera MC van Dooren M van Kempen M de Krijger R Grosveld F Tibboel D Rottier R 《American journal of physiology. Lung cellular and molecular physiology》2005,288(1):L141-L149
Although several molecular players have been described that play a role during the early phases of lung development, it is still unknown how the vasculature develops in relation to the airways. Two opposing models describe development of lung vasculature: one suggests that both vasculogenesis and angiogenesis are involved, whereas the second describes vasculogenesis as the primary mechanism. Therefore, we examined the development of the murine pulmonary vasculature through a morphological analysis from the onset of lung development [9.5 days postcoital (dpc)] until the pseudoglandular stage (13.5 dpc). We analyzed fetal lungs of Tie2-LacZ transgenic mice as well as serial sections of wild-type lungs stained with endothelial-specific antibodies (Flk-1, Fli-1, and PECAM-1). Embryos were processed with intact blood circulation to maintain the integrity of the vasculature; hence individual vessels could be identified with accuracy through serial section analysis. Furthermore, circulating primitive erythrocytes, formed exclusively by the blood islands in the yolk sac, are trapped in vessels during fixation, which proves the connection with the embryonic circulation. We report that from the first morphological sign of lung development, a clear vascular network exists that is in contact with the embryonic circulation. We propose distal angiogenesis as a new concept for early pulmonary vascular morphogenesis. In this model, capillary networks surround the terminal buds and expand by formation of new capillaries from preexisting vessels as the lung bud grows. The fact that at an early embryonic stage a complete vascular network exists may be important for the general understanding of embryonic development. 相似文献
9.
David R. Jansen Gerard C. Krijger Judith Wagener Ratanang M. Senwedi Kenneth Gabanamotse Mpho Kgadiete Zvonimir I. Kolar Jan Rijn Zeevaart 《Journal of inorganic biochemistry》2009,103(9):1265-1272
In an attempt to elucidate the in vivo stability of the prospective radiopharmaceutical [117mSn]Sn(IV)-PEI-MP, where PEI-MP stands for N,N′,N′-trimethylenephosphonate-polyethyleneimine, glass electrode potentiometry was used to determine the stability constants of the Sn4+ ion as complexed with a variety of physiological amino acids. In addition, linear free energy relationship (LFER) correlation plots were used to extrapolate the constants of the major blood plasma ligands, based on data from Cu2+, Pb2+, and Zn2+. In so doing, a thermodynamic model of blood plasma was established for Sn4+ from which the complexation tendencies of Sn4+ were predicted in the event of the intravenous administration of such a drug. It was found that the Sn(IV)-PEI-MP could succumb to competition by the glutamine amino acid, which forms more stable complex(es), whilst the PEI-MP gets taken up largely by Ca2+. Also, this study shows the value of the in vitro experiments and modeling performed for radiopharmaceutical research and for attempts to reduce the number of animal experiments. 相似文献
10.
Ana-Maria Sevcenco Gerard C. Krijger Martijn W. H. Pinkse Peter D. E. M. Verhaert Wilfred R. Hagen Peter-Leon Hagedoorn 《Journal of biological inorganic chemistry》2009,14(4):631-640
A combination of techniques to separate and quantify the native proteins associated with a particular transition metal ion
from a cellular system has been developed. The procedure involves four steps: (1) labeling of the target proteins with a suitable
short-lived radioisotope (suitable isotopes are 64Cu, 67Cu, 187W, 99Mo, 69Zn, 56Mn, 65Ni); (2) separation of intact soluble holoproteins using native isoelectric focusing combined with blue native polyacrylamide
gel electrophoresis into native–native 2D gel electrophoresis; (3) spot visualization and quantification using autoradiography;
and (4) protein identification with tandem mass spectrometry. The method was applied to the identification of copper proteins
from a soluble protein extract of wild-type Escherichia coli K12 using the radioisotope 64Cu. The E. coli protein CueO, which has previously been only identified as a multicopper oxidase following homologous overexpression, was
now directly detected as a copper protein against a wild-type background at an expression level of 0.007% of total soluble
protein. The retention of the radioisotope by the copper proteins throughout the separation process corroborates the method
to be genuinely native. The procedure developed here can be applied to cells of any origin, and to any metal having suitable
radioisotopes. The finding that the periplasmic protein CueO is the only major form of soluble protein bound copper in E. coli strengthens the view that the bacterial periplasm contains only a few periplasmic copper proteins, and that the cytosol is
devoid of copper proteins.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献