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1.
Clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated (cas) genes constitute the adaptive immune system in bacteria and archaea. Although the CRISPR-Cas systems have been hypothesized to encode potential toxins, no experimental data supporting the hypothesis are available in the literature. In this work, we provide the first experimental evidence for the presence of a toxin gene in the type I-A CRISPR system of hyperthermophilic archaeon Sulfolobus. csa5, under the control of its native promoter in a shuttle vector, could not be transformed into CRISPR-deficient mutant Sulfolobus solfataricus Sens1, demonstrating a strong toxicity in the cells. A single-amino-acid mutation destroying the intersubunit bridge of Csa5 attenuated the toxicity, indicative of the importance of Csa5 oligomerization for its toxicity. In line with the absence of Csa5 toxicity in S. solfataricus InF1 containing functional CRISPR systems, the expression of csa5 is repressed in InF1 cells. Induced from the arabinose promoter in Sens1 cells, Csa5 oligomers resistant to 1% SDS co-occur with chromosome degradation and cell death, reinforcing the connection between Csa5 oligomerization and its toxicity. Importantly, a rudivirus was shown to induce Csa5 expression and the formation of SDS-resistant Csa5 oligomers in Sulfolobus cells. This demonstrates that the derepression of csa5 and the subsequent Csa5 oligomerization take place in native virus-host systems. Thus, csa5 is likely to act as a suicide gene under certain circumstances to inhibit virus spreading. 相似文献
2.
Honglei Sun Yipeng Sun Juan Pu Yi Zhang Qingyu Zhu Jing Li Jiang Gu Kin-Chow Chang Jinhua Liu 《Journal of virology》2014,88(1):725-729
Highly pathogenic avian influenza H5N1 virus clades 2.3.4, 2.3.2, and 7 are the dominant cocirculating H5N1 viruses in poultry in China. However, humans appear to be clinically susceptible mostly to the 2.3.4 virus clade. Here, we demonstrated that A549 cells and human macrophages infected with clade 2.3.4 viruses produced significantly more viruses than those infected with the other two clades. Likewise, clade 2.3.4-infected macrophages caused the most severe cellular damage and strongest proinflammatory response. 相似文献
3.
The causes of recurrent spontaneous abortion (RSA) and fetal malformations are multifactorial and unclear in most cases. Environmental, maternal, and genetic factors have been shown to contribute to these defects. Whole-exome sequencing (WES) is widely used to detect genetic variations associated with human diseases and has recently been successfully applied to unveil genetic causes of unexplained recurrent spontaneous abortion (URSA) and fetal malformations. Here, we review the current discovery and diagnosis strategies to identify the underlying pathogenic mutations of URSA and fetal malformations using WES technology and propose to further develop WES, both to advance our understanding of these diseases and to eventually lead to targeted therapies for reproductive disorders. 相似文献
4.
Chu-Fang Chou Xiaolin Zhu Yi-Yu Lin Karen L. Gamble W. Timothy Garvey Ching-Yi Chen 《Journal of lipid research》2015,56(2):227-240
Hepatic lipid metabolism is controlled by integrated metabolic pathways. Excess accumulation of hepatic TG is a hallmark of nonalcoholic fatty liver disease, which is associated with obesity and insulin resistance. Here, we show that KH-type splicing regulatory protein (KSRP) ablation reduces hepatic TG levels and diet-induced hepatosteatosis. Expression of period 2 (Per2) is increased during the dark period, and circadian oscillations of several core clock genes are altered with a delayed phase in Ksrp−/− livers. Diurnal expression of some lipid metabolism genes is also disturbed with reduced expression of genes involved in de novo lipogenesis. Using primary hepatocytes, we demonstrate that KSRP promotes decay of Per2 mRNA through an RNA-protein interaction and show that increased Per2 expression is responsible for the phase delay in cycling of several clock genes in the absence of KSRP. Similar to Ksrp−/− livers, both expression of lipogenic genes and intracellular TG levels are also reduced in Ksrp−/− hepatocytes due to increased Per2 expression. Using heterologous mRNA reporters, we show that the AU-rich element-containing 3′ untranslated region of Per2 is responsible for KSRP-dependent mRNA decay. These findings implicate that KSRP is an important regulator of circadian expression of lipid metabolism genes in the liver likely through controlling Per2 mRNA stability. 相似文献
5.
6.
Actinidia chlorotic ringspot‐associated virus: a novel emaravirus infecting kiwifruit plants 下载免费PDF全文
Yazhou Zheng Beatriz Navarro Guoping Wang Yanxiang Wang Zuokun Yang Wenxing Xu Chenxi Zhu Liping Wang Francesco Di Serio Ni Hong 《Molecular Plant Pathology》2017,18(4):569-581
By integrating next‐generation sequencing (NGS), bioinformatics, electron microscopy and conventional molecular biology tools, a new virus infecting kiwifruit vines has been identified and characterized. Being associated with double‐membrane‐bound bodies in infected tissues and having a genome composed of RNA segments, each one containing a single open reading frame in negative polarity, this virus shows the typical features of members of the genus Emaravirus. Five genomic RNA segments were identified. Additional molecular signatures in the viral RNAs and in the proteins they encode, together with data from phylogenetic analyses, support the proposal of creating a new species in the genus Emaravirus to classify the novel virus, which is tentatively named Actinidia chlorotic ringspot‐associated virus (AcCRaV). Bioassays showed that AcCRaV is mechanically transmissible to Nicotiana benthamiana plants which, in turn, may develop chlorotic spots and ringspots. Field surveys disclosed the presence of AcCRaV in four different species of kiwifruit vines in five different provinces of central and western China, and support the association of the novel virus with symptoms of leaf chlorotic ringspots in Actinidia. Data on the molecular features of small RNAs of 21–24 nucleotides, derived from AcCRaV RNAs targeted by host RNA silencing mechanisms, are also reported, and possible molecular pathways involved in their biogenesis are discussed. 相似文献
7.
An advanced electro-active dry adhesive,which was composed of a mushroom-shaped fibrillar dry adhesive array actuated by an Ionic Polymer Metal Composite (IPMC) artificial muscle reinforced with nitrogen-doped carbon nanocages (NCNCs),was developed to imitate the actuation of a gecko's toe.The properties of the NCNC-reinforced Nafion membrane,the electromechanical properties of the NCNC-reinforced IPMC,and the related electro-active adhesion ability were investigated.The NCNCs were uniformly dispersed in the 0.1 wt% NCNC/Nafion membrane,and there was a seamless connection with no clear interface between the dry adhesive and the IPMC.Our 0.1 wt% NCNC/Nafion-IPMC actuator shows a displacement and force that are 1.6-2 times higher than those of the recast Nafion-IPMC.This is due to the increased water uptake (25.39%) and tensile strength (24.5 MPa) of the specific 3D hollow NCNC-reinforced Nafion membrane,as well as interactions between the NCNCs and the sulfonated groups of the Nafion.The NCNC/Nafion-IPMC was used to effectively actuate the mushroom-shaped dry adhesive.The normal adhesion forces were 7.85 mN,12.1 mN,and 51.7 mN at sinusoidal voltages of 1.5 V,2.5 V,and 3.5 V,respectively,at 0.1 Hz.Under the bionic leg trail,the normal and shear forces were approximately 713.5 mN (159 mN·cm-2) and 1256.6 mN (279 mN·cm-2),respectively,which satisfy the required adhesion.This new electro-active dry adhesive can be applied for active,distributed actuation and flexible grip in robots. 相似文献
8.
Wei-Qing Li Hong-Yu Yu Yi-Ming Li Xiang Wang Jin He Hong-Zhu Yan Dao-Hua Yang Xiao-Jun Wu Li-Jun Hou Hui-Min Liu Chun-Yan Xia Yi-Cheng Lu 《Biochemical and biophysical research communications》2014
Previous studies from this laboratory indicated that microRNA-21 (miR-21) contributes to chemoresistance of glioblastoma multiforme (GBM) cells to teniposide, a type II topoisomerase inhibitor. We also showed that LRRFIP1 is a target of miR-21. In this study, we found that higher baseline LRRFIP1 expression in human GBM tissue (n = 60) is associated with better prognosis upon later treatment with teniposide. Experiments in cultured U373MG cells showed enhanced toxicity of teniposide against U373MG cells transfected with a vector that resulted in LRRFIP1 overexpression (vs. cells transfected with control vector). Experiments in nude mice demonstrated better response of LRRFIP1 overexpressing xenografts to teniposide. These findings indicate that high baseline LRRFIP1 expression in GBM is associated with better response to teniposide, and encourage exploring LRRFIP1 as a target for GBM treatment. 相似文献
9.
Dan Wu Jing Liu Baiyan Wu Bo Tu Weiguo Zhu Jianyuan Luo 《Biochemical and biophysical research communications》2014
Mutations in CLN3 gene cause juvenile neuronal ceroid lipofuscinosis (JNCL or Batten disease), an early-onset neurodegenerative disorder that is characterized by the accumulation of ceroid lipofuscin within lysosomes. The function of the CLN3 protein remains unclear and is presumed to be related to Endoplasmic reticulum (ER) stress. To investigate the function of CLN3 in the ER stress signaling pathway, we measured proliferation and apoptosis in cells transfected with normal and mutant CLN3 after treatment with the ER stress inducer tunicamycin (TM). We found that overexpression of CLN3 was sufficient in conferring increased resistance to ER stress. Wild-type CLN3 protected cells from TM-induced apoptosis and increased cell proliferation. Overexpression of wild-type CLN3 enhanced expression of the ER chaperone protein, glucose-regulated protein 78 (GRP78), and reduced expression of the proapoptotic protein CCAAT/-enhancer-binding protein homologous protein (CHOP). In contrast, overexpression of mutant CLN3 or siRNA knockdown of CLN3 produced the opposite effect. Together, our data suggest that the lack of CLN3 function in cells leads to a failure of management in the response to ER stress and this may be the key deficit in JNCL that causes neuronal degeneration. 相似文献
10.
Li Wang Huiping Zhang Jihong Qian Kanqing Wang Jianxing Zhu 《Biochemical and biophysical research communications》2014