共查询到20条相似文献,搜索用时 62 毫秒
1.
Cell death can be divided into the anti-inflammatory process of apoptosis and the
pro-inflammatory process of necrosis. Necrosis, as apoptosis, is a regulated form of cell
death, and Poly-(ADP-Ribose) Polymerase-1 (PARP-1) and Receptor-Interacting Protein (RIP)
1/3 are major mediators. We previously showed that absence or inhibition of PARP-1
protects mice from nephritis, however only the male mice. We therefore hypothesized that
there is an inherent difference in the cell death program between the sexes. We show here
that in an immune-mediated nephritis model, female mice show increased apoptosis compared
to male mice. Treatment of the male mice with estrogens induced apoptosis to levels
similar to that in female mice and inhibited necrosis. Although PARP-1 was activated in
both male and female mice, PARP-1 inhibition reduced necrosis only in the male mice. We
also show that deletion of RIP-3 did not have a sex bias. We demonstrate here that male
and female mice are prone to different types of cell death. Our data also suggest that
estrogens and PARP-1 are two of the mediators of the sex-bias in cell death. We therefore
propose that targeting cell death based on sex will lead to tailored and better treatments
for each gender. 相似文献
2.
Rosanna Pescini Gobert Monique van den Eijnden Cedric Szyndralewiez Catherine Jorand-Lebrun Dominique Swinnen Linfeng Chen Corine Gillieron Fiona Pixley Pierre Juillard Patrick Gerber Caroline Johnson-L��ger Serge Halazy Montserrat Camps Agnes Bombrun Margaret Shipp Pierre-Alain Vitte Vittoria Ardissone Chiara Ferrandi Dominique Perrin Christian Rommel Rob Hooft van Huijsduijnen 《The Journal of biological chemistry》2009,284(17):11385-11395
3.
4.
5.
Nicole C. Grieder Emmanuel Caussinus David S. Parker Kenneth Cadigan Markus Affolter Stefan Luschnig 《PloS one》2008,3(9)
Background
There is increasing evidence that tissue-specific modifications of basic cellular functions play an important role in development and disease. To identify the functions of COPI coatomer-mediated membrane trafficking in Drosophila development, we were aiming to create loss-of-function mutations in the γCOP gene, which encodes a subunit of the COPI coatomer complex.Principal Findings
We found that γCOP is essential for the viability of the Drosophila embryo. In the absence of zygotic γCOP activity, embryos die late in embryogenesis and display pronounced defects in morphogenesis of the embryonic epidermis and of tracheal tubes. The coordinated cell rearrangements and cell shape changes during tracheal tube morphogenesis critically depend on apical secretion of certain proteins. Investigation of tracheal morphogenesis in γCOP loss-of-function mutants revealed that several key proteins required for tracheal morphogenesis are not properly secreted into the apical lumen. As a consequence, γCOP mutants show defects in cell rearrangements during branch elongation, in tube dilation, as well as in tube fusion. We present genetic evidence that a specific subset of the tracheal defects in γCOP mutants is due to the reduced secretion of the Zona Pellucida protein Piopio. Thus, we identified a critical target protein of COPI-dependent secretion in epithelial tube morphogenesis.Conclusions/Significance
These studies highlight the role of COPI coatomer-mediated vesicle trafficking in both general and tissue-specific secretion in a multicellular organism. Although COPI coatomer is generally required for protein secretion, we show that the phenotypic effect of γCOP mutations is surprisingly specific. Importantly, we attribute a distinct aspect of the γCOP phenotype to the effect on a specific key target protein. 相似文献6.
7.
8.
9.
Carrie L. Flood Gina P. Rodriguez Gaobin Bao Arthur H. Shockley Yoke Wah Kow Gray F. Crouse 《PLoS genetics》2015,11(3)
It is now well established that in yeast, and likely most eukaryotic organisms, initial DNA replication of the leading strand is by DNA polymerase ε and of the lagging strand by DNA polymerase δ. However, the role of Pol δ in replication of the leading strand is uncertain. In this work, we use a reporter system in Saccharomyces cerevisiae to measure mutation rates at specific base pairs in order to determine the effect of heterozygous or homozygous proofreading-defective mutants of either Pol ε or Pol δ in diploid strains. We find that wild-type Pol ε molecules cannot proofread errors created by proofreading-defective Pol ε molecules, whereas Pol δ can not only proofread errors created by proofreading-defective Pol δ molecules, but can also proofread errors created by Pol ε-defective molecules. These results suggest that any interruption in DNA synthesis on the leading strand is likely to result in completion by Pol δ and also explain the higher mutation rates observed in Pol δ-proofreading mutants compared to Pol ε-proofreading defective mutants. For strains reverting via AT→GC, TA→GC, CG→AT, and GC→AT mutations, we find in addition a strong effect of gene orientation on mutation rate in proofreading-defective strains and demonstrate that much of this orientation dependence is due to differential efficiencies of mispair elongation. We also find that a 3′-terminal 8 oxoG, unlike a 3′-terminal G, is efficiently extended opposite an A and is not subject to proofreading. Proofreading mutations have been shown to result in tumor formation in both mice and humans; the results presented here can help explain the properties exhibited by those proofreading mutants. 相似文献
10.
11.
Lili Wei Benoit Derrien Arnaud Gautier Laura Houille-Vernes Alix Boulouis Denis Saint-Marcoux Alizée Malno? Fabrice Rappaport Catherine de Vitry Olivier Vallon Yves Choquet Francis-André Wollman 《The Plant cell》2014,26(1):353-372
Starving microalgae for nitrogen sources is commonly used as a biotechnological
tool to boost storage of reduced carbon into starch granules or lipid droplets,
but the accompanying changes in bioenergetics have been little studied so far.
Here, we report that the selective depletion of Rubisco and cytochrome
b6f complex that occurs when
Chlamydomonas reinhardtii is starved for nitrogen in the
presence of acetate and under normoxic conditions is accompanied by a marked
increase in chlororespiratory enzymes, which converts the photosynthetic
thylakoid membrane into an intracellular matrix for oxidative catabolism of
reductants. Cytochrome b6f subunits
and most proteins specifically involved in their biogenesis are selectively
degraded, mainly by the FtsH and Clp chloroplast proteases. This regulated
degradation pathway does not require light, active photosynthesis, or state
transitions but is prevented when respiration is impaired or under phototrophic
conditions. We provide genetic and pharmacological evidence that NO production
from intracellular nitrite governs this degradation pathway: Addition of a NO
scavenger and of two distinct NO producers decrease and increase, respectively,
the rate of cytochrome b6f
degradation; NO-sensitive fluorescence probes, visualized by confocal
microscopy, demonstrate that nitrogen-starved cells produce NO only when the
cytochrome b6f degradation pathway
is activated. 相似文献
12.
13.
Gloria Salazar Stephanie Zlatic Branch Craige Andrew A. Peden Jan Pohl Victor Faundez 《The Journal of biological chemistry》2009,284(3):1790-1802
The Hermansky-Pudlak syndrome is a disorder affecting endosome sorting.
Disease is triggered by defects in any of 15 mouse gene products, which are
part of five distinct cytosolic molecular complexes: AP-3, homotypic fusion
and vacuole protein sorting, and BLOC-1, -2, and -3. To identify molecular
associations of these complexes, we used in vivo cross-linking
followed by purification of cross-linked AP-3 complexes and mass spectrometric
identification of associated proteins. AP-3 was co-isolated with BLOC-1,
BLOC-2, and homotypic fusion and vacuole protein sorting complex subunits;
clathrin; and phosphatidylinositol-4-kinase type II α (PI4KIIα).
We previously reported that this membrane-anchored enzyme is a regulator of
AP-3 recruitment to membranes and a cargo of AP-3 (Craige, B.,
Salazar, G., and Faundez, V. (2008) Mol. Biol.
Cell
19,1415
-1426). Using cells deficient
in different Hermansky-Pudlak syndrome complexes, we identified that BLOC-1,
but not BLOC-2 or BLOC-3, deficiencies affect PI4KIIα inclusion into
AP-3 complexes. BLOC-1, PI4KIIα, and AP-3 belong to a tripartite
complex, and down-regulation of either PI4KIIα, BLOC-1, or AP-3
complexes led to similar LAMP1 phenotypes. Our analysis indicates that BLOC-1
complex modulates the association of PI4KIIα with AP-3. These results
suggest that AP-3 and BLOC-1 act, either in concert or sequentially, to
specify sorting of PI4KIIα along the endocytic route.Membranous organelles along the exocytic and endocytic pathways are each
defined by unique lipid and protein composition. Vesicle carriers communicate
and maintain the composition of these organelles
(2). Consequently defining the
machineries that specify vesicle formation, composition, and delivery are
central to understanding membrane protein traffic. Generally vesicle
biogenesis uses multiprotein cytosolic machineries to select membrane
components for inclusion in nascent vesicles
(2,
3). Heterotetrameric adaptor
complexes (AP-1 to AP-4) are critical to generate vesicles of specific
composition from the different organelles constituting the exocytic and
endocytic routes
(2-4).The best understood vesicle formation machinery in mammalian cells is the
one organized around the adaptor complex AP-2
(5). This complex generates
vesicles from the plasma membrane using clathrin. Our present detailed
understanding of AP-2 vesicle biogenesis mechanisms and interactions emerged
from a combination of organellar and in vitro binding proteomics
analyses together with the study of binary interactions in cell-free systems
(5-9).
In contrast, the vesicle biogenesis pathways controlled by AP-3 are far less
understood. AP-3 functions to produce vesicles that traffic selected membrane
proteins from endosomes to lysosomes, lysosome-related organelles, or synaptic
vesicles
(10-13).
AP-3 is one of the protein complexes affected in the Hermansky-Pudlak syndrome
(HPS;3 Online
Mendelian Inheritance in Man (OMIM) 203300). So far, mutations in any of 15
mouse or eight human genes trigger a common syndrome. This syndrome
encompasses defects that include pigment dilution, platelet dysfunction,
pulmonary fibrosis, and occasionally neurological phenotypes
(14,
15). All forms of HPS show
defective vesicular biogenesis or trafficking that affects lysosomes,
lysosome-related organelles (for example melanosomes and platelet dense
granules), and, in some of them, synaptic vesicles
(11-13).
Most of the 15 HPS loci encode polypeptides that assemble into five distinct
molecular complexes: the adaptor complex AP-3, HOPS, and the BLOC complexes 1,
2, and 3 (14). Recently binary
interactions between AP-3 and BLOC-1 or BLOC-1 and BLOC-2 suggested that
arrangements of these complexes could regulate membrane protein targeting
(16). Despite the abundance of
genetic deficiencies leading to HPS and genetic evidence that HPS complexes
may act on the same pathway in defined cell types
(17), we have only a partial
picture of protein interactions organizing these complexes and how they might
control membrane protein targeting.In this study, we took advantage of cell-permeant and reversible
cross-linking of HPS complexes followed by their immunoaffinity purification
to identify novel molecular interactions. Cross-linked AP-3 co-purified with
BLOC-1, BLOC-2, HOPS, clathrin, and the membrane protein PI4KIIα. We
previously identified PI4KIIα as a cargo and regulator of AP-3
recruitment to endosomes (1,
18). Using mutant cells
deficient in either individual HPS complexes or a combination of them, we
found that BLOC-1 facilitates the interaction of AP-3 and PI4KIIα. Our
studies demonstrate that subunits of four of the five HPS complexes co-isolate
with AP-3. Moreover BLOC-1, PI4KIIα, and AP-3 form a tripartite complex
as demonstrated by sequential co-immunoprecipitations as well as by similar
LAMP1 distribution phenotypes induced by down-regulation of components of this
tripartite complex. Our findings indicate that BLOC-1 complex modulates the
recognition of PI4KIIα by AP-3. These data suggest that AP-3, either in
concert or sequentially with BLOC-1, participates in the sorting of common
membrane proteins along the endocytic route. 相似文献
14.
15.
Candida antarctica lipase B (CALB) is one of the most widely used and studied enzymes in the world. In order to achieve the high-level expression of CALB in Pichia, we optimized the codons of CALB gene and α-factor by using a de novo design and synthesis strategy. Through comparative analysis of a series of recombinants with different expression components, we found that the methanol-inducible expression recombinant carrying the codon-optimized α-factor and mature CALB gene (pPIC9KαM-CalBM) has the highest lipase production capacity. After fermentation parameters optimization, the lipase activity and protein content of the recombinant pPIC9KαM-CalBM reached 6,100 U/mL and 3.0 g/L, respectively, in a 5-L fermentor. We believe this strategy could be of special interest due to its capacity to improve the expression level of target gene, and the Pichia transformants carrying the codon-optimized gene had great potential for the industrial-scale production of CALB lipase. 相似文献
16.
Girish V. Shah Anbalagan Muralidharan Mitan Gokulgandhi Kamal Soan Shibu Thomas 《The Journal of biological chemistry》2009,284(2):1018-1030
Calcitonin, a neuroendocrine peptide, and its receptor are localized in the
basal epithelium of benign prostate but in the secretory epithelium of
malignant prostates. The abundance of calcitonin and calcitonin receptor mRNA
displays positive correlation with the Gleason grade of primary prostate
cancers. Moreover, calcitonin increases tumorigenicity and invasiveness of
multiple prostate cancer cell lines by cyclic AMP-dependent protein
kinase-mediated actions. These actions include increased secretion of matrix
metalloproteinases and urokinase-type plasminogen activator and an increase in
prostate cancer cell invasion. Activation of calcitonin-calcitonin receptor
autocrine loop in prostate cancer cell lines led to the loss of cell-cell
adhesion, destabilization of tight and adherens junctions, and internalization
of key integral membrane proteins. In addition, the activation of
calcitonin-calcitonin receptor axis induced epithelial-mesenchymal transition
of prostate cancer cells as characterized by cadherin switch and the
expression of the mesenchymal marker, vimentin. The activated calcitonin
receptor phosphorylated glycogen synthase kinase-3, a key regulator of
cytosolic β-catenin degradation within the WNT signaling pathway. This
resulted in the accumulation of intracellular β-catenin, its
translocation in the nucleus, and transactivation of β-catenin-responsive
genes. These results for the first time identify actions of
calcitonin-calcitonin receptor axis on prostate cancer cells that lead to the
destabilization of cell-cell junctions, epithelial-to-mesenchymal transition,
and activation of WNT/β-catenin signaling. The results also suggest that
cyclic AMP-dependent protein kinase plays a key role in calcitonin
receptor-induced destabilization of cell-cell junctions and activation of
WNT-β-catenin signaling.Prostate cancer
(PC)2 is the most
commonly diagnosed cancer and the second leading cause of cancer deaths in men
in the United States (1,
2). Although androgen ablation
therapy is effective in men with advanced disease for some time, the disease
subsequently progresses to the androgen-independent stage. The population of
prostate cells expressing neuroendocrine factors such as calcitonin (CT) also
increases during this progression
(3–5).
At this stage, the disease is metastatic and chemoresistant. Present evidence
suggests that cancer metastasis is usually preceded by the disruption of
normal cell-cell adhesion and the loss of integrity of the primary tumor site
(6,
7). This process may include
several genetic, molecular, and morphological changes characterized by
epithelial-to-mesenchymal transition (EMT)
(8–10).
The EMT is characterized by the loss of cell polarity, altered cell-cell and
cell-matrix adhesion, and acquisition of migratory, mesenchymal phenotype.
Other reported changes include down-regulation of E-cadherin, induction of
N-cadherin, release of β-catenin from junctional complexes, and its
translocation to the nucleus
(11–13).
However, the precise molecular mechanisms associated with this process are
obscure.Several growth factors, including hepatocyte growth factor, transforming
growth factor-β, vascular endothelial growth factor, and epidermal growth
factor, have been reported to induce EMT in tumor cell lines
(14–16).
We have shown that the expression of CT and its G protein-coupled receptor
(CTR) is remarkably higher in advanced PCs, and the CT-CTR autocrine axis is a
potent stimulator of PC cell tumorigenicity, invasion, and metastasis
(4,
17–19).
Although CT-stimulated increase in the motility and invasion of PC cells may
be mediated by CT-stimulated secretion of matrix metalloproteinases and
urokinase-type plasminogen activator, the precise molecular mechanisms
preceding these CTR actions remain to be elucidated
(18,
20). We tested the hypothesis
that CT induces biochemical and morphological changes associated with EMT to
increase the invasiveness of PC cells.Our results indicate that activation of the CT-CTR autocrine axis in
prostate cancer cells induced several changes associated with EMT such as
remodeling of tight and adherens junctions, cadherin switching, and activation
of WNT/β-catenin signaling. In contrast, the silencing of the CT-CTR axis
reversed this process. Moreover, cyclic AMP-dependent protein kinase (PKA)
plays a key role in this CT-CTR-mediated process. This is the first study
demonstrating the action of prostate CTR on junctional complexes and
WNT/β-catenin signaling of PC cell lines. 相似文献
17.
18.
19.
Toru Yoshihara Kazushi Sugihara Yasuhiko Kizuka Shogo Oka Masahide Asano 《The Journal of biological chemistry》2009,284(18):12550-12561
The glycosylation of glycoproteins and glycolipids is important for central
nervous system development and function. Although the roles of several
carbohydrate epitopes in the central nervous system, including polysialic
acid, the human natural killer-1 (HNK-1) carbohydrate, α2,3-sialic acid,
and oligomannosides, have been investigated, those of the glycan backbone
structures, such as Galβ1-4GlcNAc and Galβ1-3GlcNAc, are not fully
examined. Here we report the generation of mice deficient in
β4-galactosyltransferase-II (β4GalT-II). This galactosyltransferase
transfers Gal from UDP-Gal to a nonreducing terminal GlcNAc to synthesize the
Gal β1-4GlcNAc structure, and it is strongly expressed in the central
nervous system. In behavioral tests, the β4GalT-II-/- mice
showed normal spontaneous activity in a novel environment, but impaired
spatial learning/memory and motor coordination/learning. Immunohistochemistry
showed that the amount of HNK-1 carbohydrate was markedly decreased in the
brain of β4GalT-II-/- mice, whereas the expression of
polysialic acid was not affected. Furthermore, mice deficient in
glucuronyltransferase (GlcAT-P), which is responsible for the biosynthesis of
the HNK-1 carbohydrate, also showed impaired spatial learning/memory as
described in our previous report, although their motor coordination/learning
was normal as shown in this study. Histological examination showed abnormal
alignment and reduced number of Purkinje cells in the cerebellum of
β4GalT-II-/- mice. These results suggest that the
Galβ1-4GlcNAc structure in the HNK-1 carbohydrate is mainly synthesized
by β4GalT-II and that the glycans synthesized by β4GalT-II have
essential roles in higher brain functions, including some that are
HNK-1-dependent and some that are not.The glycosylation of glycoproteins, proteoglycans, and glycolipids is
important for their biological activities, stability, transport, and clearance
from circulation, and cell-surface glycans participate in cell-cell and
cell-extracellular matrix interactions. In the central nervous system, several
specific carbohydrate epitopes, including polysialic acid
(PSA),3 the
human natural killer-1 (HNK-1) carbohydrate, α2,3-sialic acid, and
oligomannosides play indispensable roles in neuronal generation, cell
migration, axonal outgrowth, and synaptic plasticity
(1). Functional analyses of the
glycan backbone structures, like lactosamine core (Galβ1-4GlcNAc),
neolactosamine core (Galβ1-3GlcNAc), and polylactosamine
(Galβ1-4GlcNAcβ1-3) have been carried out using gene-deficient mice
in β4-galactosyltransferase-I (β4GalT-I)
(2,
3), β4GalT-V
(4),
β3-N-acetylglucosaminyl-transferase-II (β3GnT-II)
(5), β3GnT-III
(Core1-β3GnT) (6),
β3GnT-V (7), and Core2GnT
(8). However, the roles of
these glycan backbone structures in the nervous system have not been examined
except the olfactory sensory system
(9).β4GalTs synthesize the Galβ1-4GlcNAc structure via the
β4-galactosylation of glycoproteins and glycolipids; the β4GalTs
transfer galactose (Gal) from UDP-Gal to a nonreducing terminal
N-acetylglucosamine (GlcNAc) of N- and O-glycans
with a β-1,4-linkage. The β4GalT family has seven members
(β4GalT-I to VII), of which at least five have similar
Galβ1-4GlcNAc-synthesizing activities
(10,
11). Each β4GalT has a
tissue-specific expression pattern and substrate specificity with overlapping,
suggesting each β4GalT has its own biological role as well as redundant
functions. β4GalT-I and β4GalT-II share the highest identity (52% at
the amino acid level) among the β4GalTs
(12), suggesting these two
galactosyltransferases can compensate for each other. β4GalT-I is
strongly and ubiquitously expressed in various non-neural tissues, whereas
β4GalT-II is strongly expressed in neural tissues
(13,
14). Indeed, the β4GalT
activity in the brain of β4GalT-I-deficient (β4GalT-I-/-)
mice remains as high as 65% of that of wild-type mice, and the expression
levels of PSA and the HNK-1 carbohydrate in the brain of these mice are normal
(15). These results suggest
β4GalTs other than β4GalT-I, like β4GalT-II, are important in
the nervous system.Among the β4GalT family members, only β4GalT-I-/- mice
have been examined extensively; this was done by us and another group. We
reported that glycans synthesized by β4GalT-I play various roles in
epithelial cell growth and differentiation, inflammatory responses, skin wound
healing, and IgA nephropathy development
(2,
16-18).
Another group reported that glycans synthesized by β4GalT-I are involved
in anterior pituitary hormone function and in fertilization
(3,
19). However, no other nervous
system deficits have been reported in these mice, and the role of the
β4-galactosylation of glycoproteins and glycolipids in the nervous system
has not been fully examined.In this study, we generated β4GalT-II-/- mice and examined
them for behavioral abnormalities and biochemical and histological changes in
the central nervous system. β4GalT-II-/- mice were impaired in
spatial learning/memory and motor coordination/learning. The amount of HNK-1
carbohydrate was markedly decreased in the β4GalT-II-/- brain,
but PSA expression was not affected. These results suggest that the
Galβ1-4GlcNAc structure in the HNK-1 carbohydrate is mainly synthesized
by β4GalT-II and that glycans synthesized by β4GalT-II have
essential roles in higher brain functions, including ones that are HNK-1
carbohydrate-dependent and ones that are independent of HNK-1. 相似文献
20.
Isolation of the Ornithine-δ-Aminotransferase cDNA and Effect
of Salt Stress on Its Expression in Arabidopsis
thaliana
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Nancy H.C.J. Roosens Tran T. Thu Hayati M. Iskandar Michel Jacobs 《Plant physiology》1998,117(1):263-271
To evaluate the relative importance of ornithine (Orn) as a precursor in proline (Pro) synthesis, we isolated and sequenced a cDNA encoding the Orn-δ-aminotransferase (δ-OAT) from Arabidopsis thaliana. The deduced amino acid sequence showed high homology with bacterial, yeast, mammalian, and plant sequences, and the N-terminal residues exhibited several common features with a mitochondrial transit peptide. Our results show that under both salt stress and normal conditions, δ-OAT activity and mRNA in young plantlets are slightly higher than in older plants. This appears to be related to the necessity to dispose of an easy recycling product, glutamate. Analysis of the expression of the gene revealed a close association with salt stress and Pro production. In young plantlets, free Pro content, Δ1-pyrroline-5-carboxylate synthase mRNA, δ-OAT activity, and δ-OAT mRNA were all increased by salt-stress treatment. These results suggest that for A. thaliana, the Orn pathway, together with the glutamate pathway, plays an important role in Pro accumulation during osmotic stress. Conversely, in 4-week-old A. thaliana plants, although free Pro level also increased under salt-stress conditions, the δ-OAT activity appeared to be unchanged and δ-OAT mRNA was not detectable. Δ1-pyrroline-5-carboxylate synthase mRNA was still induced at a similar level. Therefore, for the adult plants the free Pro increase seemed to be due to the activity of the enzymes of the glutamate pathway. 相似文献