iRhom2 regulation of TACE controls TNF-mediated protection against Listeria and responses to LPS

Innate immune responses are vital for pathogen defense but can result in septic shock when excessive. A key mediator of septic shock is tumor necrosis factor-α (TNFα), which is shed from the plasma membrane after cleavage by the TNFα convertase (TACE). We report that the rhomboid family member iRhom2 interacted with TACE and regulated TNFα shedding. iRhom2 was critical for TACE maturation and trafficking to the cell surface in hematopoietic cells. Gene-targeted iRhom2-deficient mice showed reduced serum TNFα in response to lipopolysaccharide (LPS) and could survive a lethal LPS dose. Furthermore, iRhom2-deficient mice failed to control the replication of Listeria monocytogenes. Our study has identified iRhom2 as a regulator of innate immunity that may be an important target for modulating sepsis and pathogen defense.     

胆固醇平衡调控中的SREBP途径

胆固醇在生物体内发挥着极其重要的作用。细胞主要通过调节胆固醇的合成、吸收、酯化及外流等途径之间的平衡以维持正常的胆固醇浓度。本文介绍甾醇调控元件结合蛋白(SREBP,Sterolregulatoryelement-bindingproteins)转录因子对胆固醇平衡的调控。SREBP主要调节胆固醇的合成、LDL受体介导的胆固醇的吸收和脂肪酸的合成。SREBP前体是一内质网膜蛋白,SREBP经两次蛋白酶水解后释放出其N端结构域,进入细胞核,激活靶基因的转录。随着对SREBP领域研究的深入,人们对胆固醇平衡调节将认识更多,并有可能开发出更为有效的控制胆固醇的药物。     

ER cargo properties specify a requirement for COPII coat rigidity mediated by Sec13p

Eukaryotic secretory proteins exit the endoplasmic reticulum (ER) via transport vesicles generated by the essential coat protein complex II (COPII) proteins. The outer coat complex, Sec13-Sec31, forms a scaffold that is thought to enforce curvature. By exploiting yeast bypass-of-sec-thirteen (bst) mutants, where Sec13p is dispensable, we probed the relationship between a compromised COPII coat and the cellular context in which it could still function. Genetic and biochemical analyses suggested that Sec13p was required to generate vesicles from membranes that contained asymmetrically distributed cargoes that were likely to confer opposing curvature. Thus, Sec13p may rigidify the COPII cage and increase its membrane-bending capacity; this function could be bypassed when a bst mutation renders the membrane more deformable.     

Distinct modes of signal recognition particle interaction with the ribosome

Signal recognition particle (SRP), together with its receptor (SR), mediates the targeting of ribosome-nascent chain complexes to the endoplasmic reticulum. Using protein cross-linking, we detected distinct modes in the binding of SRP to the ribosome. During signal peptide recognition, SRP54 is positioned at the exit site close to ribosomal proteins L23a and L35. When SRP54 contacts SR, SRP54 is rearranged such that it is no longer close to L23a. This repositioning may allow the translocon to dock with the ribosome, leading to insertion of the signal peptide into the translocation channel.     

Chaperone activity of protein O-fucosyltransferase 1 promotes notch receptor folding

Notch proteins are receptors for a conserved signaling pathway that affects numerous cell fate decisions. We found that in Drosophila, Protein O-fucosyltransferase 1 (OFUT1), an enzyme that glycosylates epidermal growth factor-like domains of Notch, also has a distinct Notch chaperone activity. OFUT1 is an endoplasmic reticulum protein, and its localization was essential for function in vivo. OFUT1 could bind to Notch, was required for the trafficking of wild-type Notch out of the endoplasmic reticulum, and could partially rescue defects in secretion and ligand binding associated with Notch point mutations. This ability of OFUT1 to facilitate folding of Notch did not require its fucosyltransferase activity. Thus, a glycosyltransferase can bind its substrate in the endoplasmic reticulum to facilitate normal folding.     

葡萄糖调节蛋白78研究进展

葡萄糖调节蛋白78(Glucose regulated protein 78ku, GRP78)又称免疫球蛋白重链结合蛋白(Immunoglobulin heavy chain binding protein, Bip),是位于内质网上的一种重要分子伴侣,属热休克蛋白70家族的一员,GRP78分子及其DNA分子序列结构在许多生物物种中高度保守.GRP78在内质网中参与阻止内质网新生肽聚集、调节内质网钙稳态、抗内质网相关性细胞凋亡以及启动未折叠蛋白反应等.近年来发现,GRP78与多种疾病发生发展密切相关,GRP78生物学功能研究已经引起广泛关注.     

Requirement of GTP hydrolysis for dissociation of the signal recognition particle from its receptor.

The signal recognition particle (SRP) directs signal sequence specific targeting of ribosomes to the rough endoplasmic reticulum. Displacement of the SRP from the signal sequence of a nascent polypeptide is a guanosine triphosphate (GTP)-dependent reaction mediated by the membrane-bound SRP receptor. A nonhydrolyzable GTP analog can replace GTP in the signal sequence displacement reaction, but the SRP then fails to dissociate from the membrane. Complexes of the SRP with its receptor containing the nonhydrolyzable analog are incompetent for subsequent rounds of protein translocation. Thus, vectorial targeting of ribosomes to the endoplasmic reticulum is controlled by a GTP hydrolysis cycle that regulates the affinity between the SRP, signal sequences, and the SRP receptor.     

Antigen presentation requires transport of MHC class I molecules from the endoplasmic reticulum

The role of exocytosis of major histocompatibility complex (MHC) class I molecules in the presentation of antigens to mouse cytotoxic T lymphocytes (CTLs) was examined by use of a recombinant vaccinia virus that expresses the E19 glycoprotein from adenovirus. E19 blocked the presentation of vaccinia and influenza virus proteins to CTLs in a MHC class I allele-specific manner identical to its inhibition of MHC class I transport from the endoplasmic reticulum. This finding indicates that (i) the relevant parameter for antigen presentation is the rate of MHC class I molecule exocytosis, not the level of class I cell surface expression, and (ii) association of class I molecules with antigen is likely to occur within the endoplasmic reticulum.     

Coordinated activation of Hsp70 chaperones

Hsp70s are a ubiquitous family of molecular chaperones involved in many cellular processes. Two Hsp70s, Lhs1p and Kar2p, are required for protein biogenesis in the yeast endoplasmic reticulum. Here, we found that Lhs1p and Kar2p specifically interacted to couple, and coordinately regulate, their respective activities. Lhs1p stimulated Kar2p by providing a specific nucleotide exchange activity, whereas Kar2p reciprocally activated the Lhs1p adenosine triphosphatase (ATPase). The two ATPase activities are coupled, and their coordinated regulation is essential for normal function in vivo.     

Proapoptotic BAX and BAK modulate the unfolded protein response by a direct interaction with IRE1alpha

Accumulation of misfolded protein in the endoplasmic reticulum (ER) triggers an adaptive stress response-termed the unfolded protein response (UPR)-mediated by the ER transmembrane protein kinase and endoribonuclease inositol-requiring enzyme-1alpha (IRE1alpha). We investigated UPR signaling events in mice in the absence of the proapoptotic BCL-2 family members BAX and BAK [double knockout (DKO)]. DKO mice responded abnormally to tunicamycin-induced ER stress in the liver, with extensive tissue damage and decreased expression of the IRE1 substrate X-box-binding protein 1 and its target genes. ER-stressed DKO cells showed deficient IRE1alpha signaling. BAX and BAK formed a protein complex with the cytosolic domain of IRE1alpha that was essential for IRE1alpha activation. Thus, BAX and BAK function at the ER membrane to activate IRE1alpha signaling and to provide a physical link between members of the core apoptotic pathway and the UPR.     

Defective presentation of endogenous antigen by a cell line expressing class I molecules

Cytotoxic T lymphocytes (CTLs) recognize class I major histocompatibility complex (MHC) molecules associated with antigenic peptides derived from endogenously synthesized proteins. Binding to such peptides is a requirement for class I assembly in the endoplasmic reticulum (ER). A mutant human cell line, T2, assembles and transports to its surface some, but not all, class I MHC molecules. The class I molecules expressed on the surface of T2 do not present peptides derived from cytosolic antigens, although they can present exogenously added peptides to CTL. The transported class I molecules may interact weakly with an unknown retaining factor in the ER such that they can assemble despite the relative shortage of peptides.     

Golgin tethers define subpopulations of COPI vesicles

Coiled-coil proteins of the golgin family have been implicated in intra-Golgi transport through tethering coat protein complex I (COPI) vesicles. The p115-golgin tether is the best studied, and here we characterize the golgin-84-CASP tether. The vesicles bound by this tether were strikingly different from those bound by the p115-golgin tether in that they lacked members of the p24 family of putative cargo receptors and contained enzymes instead of anterograde cargo. Microinjected golgin-84 or CASP also inhibited Golgi-enzyme transport to the endoplasmic reticulum, further implicating this tether in retrograde transport. These and other golgins may modulate the flow patterns within the Golgi stack.     

Diurnal rhythm in endoplasmic reticulum of rat liver: electron microscopic study

Electron microscopy of rat hepatocytes revealed a diurnal variation in the relative amounts of endoplasmic reticulum structures and regional differences in their distribution within the hepatic lobule. The diurnal changes in smooth and rough endoplasmic reticulum structures were compared with the diurnal changes in the hepatic microsomal enzyme hexobarbital oxidase. In the control group, at the time when enzyme activity was maximum, the amount of smooth endoplasmic reticulum was also maximum and vice versa. When the enzyme rhythm was abolished, as in blinded rats, the diurnal rhythm in the endoplasmic reticulum was also abolished.     

S351—1西瓜雄性不育的超微结构研究

在西瓜Ｓ３５１－１雄性不育材料遗传学及细胞分析的基础上，对其超微结构进行了观察，结果表明，次级造孢细胞在败育过程中，除细胞质壁分离、核解体消失外，细胞质中细胞器的形态与结构亦表现异常，尤其是溶酶体及液泡的数目与形态的变化。解体核周期细胞质中长丝状或封闭环状的内质网复合结构出现等，并对次级造孢细胞中液泡系的动态、内质网的结构异常与败育的关系进行了讨论。     

Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death

Multiple death signals influence mitochondria during apoptosis, yet the critical initiating event for mitochondrial dysfunction in vivo has been unclear. tBID, the caspase-activated form of a "BH3-domain-only" BCL-2 family member, triggers the homooligomerization of "multidomain" conserved proapoptotic family members BAK or BAX, resulting in the release of cytochrome c from mitochondria. We find that cells lacking both Bax and Bak, but not cells lacking only one of these components, are completely resistant to tBID-induced cytochrome c release and apoptosis. Moreover, doubly deficient cells are resistant to multiple apoptotic stimuli that act through disruption of mitochondrial function: staurosporine, ultraviolet radiation, growth factor deprivation, etoposide, and the endoplasmic reticulum stress stimuli thapsigargin and tunicamycin. Thus, activation of a "multidomain" proapoptotic member, BAX or BAK, appears to be an essential gateway to mitochondrial dysfunction required for cell death in response to diverse stimuli.     

运动、胸腺细胞凋亡与机制研究

主要通过文献资料法,对细胞凋亡的机制及运动与胸腺细胞凋亡的关系等进行了述评,其中细胞凋亡与死亡受体信号转导途径、细胞凋亡与线粒体凋亡途径和细胞凋亡与内质网凋亡途径是本文论述的重点。结果显示,有关胸腺细胞凋亡的死亡受体信号转导途径机制和线粒体机制研究不多,有关胸腺细胞凋亡的内质网机制甚少。现有研究显示运动诱导胸腺细胞凋亡与线粒体凋亡途径关系密切,其他有待进一步研究证实。     

牛卵母细胞发育的超微结构研究

本实验根据卵细胞的数量及分布、透明带的变化,卵泡大小等把牛卵母细胞的整个发育过程分为六个阶段,在超微结构的水平上进行了研究。随着卵母细胞的发育,高尔基体、线粒体、滑面内质网、皮质颗粒等细胞器数量不断增加,且逐渐迁移到皮质区。卵母细胞成熟时。高尔基体及粗面内质网消失,皮质颗粒开始在质膜下呈线状排列,而线粒体又向胞质中央区扩散。直径2毫米以上卵泡卵母细胞中,少嵴圆形线粒体一律转变为带帽线粒体,核仁发生致密化。此外,牛卵母细胞中有带有界膜的、均匀分布的囊泡。皮质颗粒成团发生和分布也是牛卵母细胞的特点之一。实验事实证明,皮质颗粒的形成与滑面内质网关系更为密切。     

Peptide binding and release by proteins implicated as catalysts of protein assembly

Two members of the hsp70 family, termed hsc70 and BiP, have been implicated in promoting protein folding and assembly processes in the cytoplasm and the lumen of the endoplasmic reticulum, respectively. Short hydrophilic (8 to 25 residues) synthetic peptides have now been tested as possible mimics of polypeptide chain substrates to help define an enzymatic basis for these activities. Both BiP and hsc70 have specific peptide binding sites. Peptide binding elicits hydrolysis of adenosine triphosphate, with the subsequent release of bound peptide.