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https://www.readbyqxmd.com/read/29161591/cell-invasion-in%C3%A2-vivo-via-rapid-exocytosis-of-a-transient-lysosome-derived-membrane-domain
#1
Kaleb M Naegeli, Eric Hastie, Aastha Garde, Zheng Wang, Daniel P Keeley, Kacy L Gordon, Ariel M Pani, Laura C Kelley, Meghan A Morrissey, Qiuyi Chi, Bob Goldstein, David R Sherwood
Invasive cells use small invadopodia to breach basement membrane (BM), a dense matrix that encases tissues. Following the breach, a large protrusion forms to clear a path for tissue entry by poorly understood mechanisms. Using RNAi screening for defects in Caenorhabditis elegans anchor cell (AC) invasion, we found that UNC-6(netrin)/UNC-40(DCC) signaling at the BM breach site directs exocytosis of lysosomes using the exocyst and SNARE SNAP-29 to form a large protrusion that invades vulval tissue. Live-cell imaging revealed that the protrusion is enriched in the matrix metalloprotease ZMP-1 and transiently expands AC volume by more than 20%, displacing surrounding BM and vulval epithelium...
November 20, 2017: Developmental Cell
https://www.readbyqxmd.com/read/29160905/regulated-recruitment-of-srgap1-modulates-rhoa-signaling-for-contractility-during-epithelial-junction-maturation
#2
Xuan Liang, Sajini Kiru, Guillermo A Gomez, Alpha S Yap
Adherens junctions in epithelia are contractile structures, where coupling of adhesion to the actomyosin cytoskeleton generates mechanical tension for morphogenesis and homeostasis. In established monolayers, junctional contractility is supported by the interplay between cell signals and scaffolding proteins. However, less is known about how contractile junctions develop, especially during the establishment of epithelial monolayers. Here we show that junctional tension increases concomitant with accumulation of actomyosin networks as Caco-2 epithelia become confluent...
November 21, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/29158625/comparative-genomic-hybridization-and-transcriptome-sequencing-reveal-that-two-genes-osi_14279-loc_os03g62620-and-osi_10794-loc_os03g14950-regulate-the-mutation-in-the-%C3%AE-rl-rice-mutant
#3
Xulong Wang, Fanhua Wang, Huiqiong Chen, Xiaoyu Liang, Yingmei Huang, Jicai Yi
We previously established the genetic locus of the rolled-leaf mutant, γ-rl, to chromosome 3. In this study, we performed a comparative genomic hybridization (CGH) analysis to identify the genes responsible for the γ-rl mutant phenotype. This was combined with RNA transcriptome sequencing (RNA-seq) to analyze differences in the mRNA expression in seeds 12 h after germination. Using the reference genome of the "indica type" rice from GenBank, we created a chip with 386,000 high density DNA probes designed to target chromosome 3...
October 2017: Physiology and Molecular Biology of Plants: An International Journal of Functional Plant Biology
https://www.readbyqxmd.com/read/29158600/crispr-cas9-library-screening-for-drug-target-discovery
#4
REVIEW
Morito Kurata, Kouhei Yamamoto, Branden S Moriarity, Masanobu Kitagawa, David A Largaespada
CRISPR/Cas9-based tools have rapidly developed in recent years. These include CRISPR-based gene activation (CRISPRa) or inhibition (CRISPRi), for which there are libraries. CRISPR libraries for loss of function have been widely used to identify new biological mechanisms, such as drug resistance and cell survival signals. CRISPRa is highly useful in screening for gain of functions, and CRISPRi is a more powerful tool than RNA interference (RNAi) libraries in screening for loss of functions. Positive selection using a CRISPR library can detect survival cells with specific conditions, such as drug treatment, and it can easily clarify drug resistance mechanisms...
November 20, 2017: Journal of Human Genetics
https://www.readbyqxmd.com/read/29158292/sphingomyelin-metabolism-is-a-regulator-of-kras-function
#5
Dharini van der Hoeven, Kwang-Jin Cho, Yong Zhou, Xiaoping Ma, Wei Chen, Ali Naji, Dina Montufar-Solis, Yan Zuo, Sarah E Kovar, Kandice R Levental, Jeffrey A Frost, Ransome van der Hoeven, John F Hancock
KRAS must localize to the plasma membrane (PM) for biological activity. We show here that multiple acid sphingomyelinase (ASM) inhibitors, including tricyclic antidepressants, mislocalized phosphatidylserine (PtdSer) and KRASG12V from the PM; resulting in abrogation of KRASG12V signaling and potent, selective growth inhibition of mutant KRAS transformed cancer cells. Concordantly, in nude mice, the ASM inhibitor fendiline decreased the rate of growth of oncogenic KRAS-expressing MiaPaCa-2 tumors, but had no effect on the growth of the wild-type KRAS-expressing BxPC-3 tumors...
November 20, 2017: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/29158205/a-molting-inhibiting-hormone-like-protein-from-pacific-white-shrimp-litopenaeus-vannamei-is-involved-in-immune-responses
#6
Hongliang Zuo, Jia Yuan, Shengwen Niu, Linwei Yang, Shaoping Weng, Jianguo He, Xiaopeng Xu
The molting-inhibiting hormones (MIHs) from the crustacean hyperglycemic hormone (CHH) family are a group of neuropeptides that are implicated in regulation of molting and reproduction in crustaceans. In this study, a novel protein containing a typical crustacean neuropeptide domain was identified from Litopenaeus vannamei. The protein showed high homology with other shrimp MIHs and was then designated as a MIH-like protein (MIHL). Among the detected tissues, the heart expressed the highest level of MIHL. The expression of MIHL could be significantly up-regulated after infection with white spot syndrome virus (WSSV), gram-negative bacterium Vibro parahaemolyticus and gram-positive bacterium Staphylococcus aureus, indicating that MIHL could be involved in immune responses...
November 17, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29157132/in-planta-functions-of-cytochrome-p450-monooxygenase-genes-in-the-phytocassane-biosynthetic-gene-cluster-on-rice-chromosome-2
#7
Zhongfeng Ye, Kohei Yamazaki, Hiromi Minoda, Koji Miyamoto, Sho Miyazaki, Hiroshi Kawaide, Arata Yajima, Hideaki Nojiri, Hisakazu Yamane, Kazunori Okada
In response to environmental stressors such as blast fungal infections, rice produces phytoalexins, an antimicrobial diterpenoid compound. Together with momilactones, phytocassanes are among the major diterpenoid phytoalexins. The biosynthetic genes of diterpenoid phytoalexin are organized on the chromosome in functional gene clusters, comprising diterpene cyclase, dehydrogenase, and cytochrome P450 monooxygenase genes. Their functions have been studied extensively using in vitro enzyme assay systems. Specifically, P450 genes (CYP71Z6, Z7; CYP76M5, M6, M7, M8) on rice chromosome 2 have multifunctional activities associated with ent-copalyl diphosphate-related diterpene hydrocarbons, but the in planta contribution of these genes to diterpenoid phytoalexin production remains unknown...
November 20, 2017: Bioscience, Biotechnology, and Biochemistry
https://www.readbyqxmd.com/read/29156673/induction-of-dise-in-ovarian-cancer-cells-in-vivo
#8
Andrea E Murmann, Kaylin M McMahon, Ashley Haluck-Kangas, Nandini Ravindran, Monal Patel, Calvin Y Law, Sonia Brockway, Jian-Jun Wei, C Shad Thaxton, Marcus E Peter
The death receptor CD95/Fas can be activated by immune cells to kill cancer cells. shRNAs and siRNAs derived from CD95 or CD95 ligand (CD95L) are highly toxic to most cancer cells. We recently found that these sh/siRNAs kill cancer cells in the absence of the target by targeting the 3'UTRs of critical survival genes through canonical RNAi. We have named this unique form of off-target effect DISE (for death induced by survival gene elimination). DISE preferentially kills transformed cells and cancer stem cells...
October 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/29155327/expression-profiles-of-the-p38-mapk-signaling-pathway-from-chinese-shrimp-fenneropenaeus-chinensis-in-response-to-viral-and-bacterial-infections
#9
Yuying He, Wanlong Yao, Ping Liu, Jian Li, Qingyin Wang
Mitogen-activated protein kinase (MAPK) signaling pathway plays an important role in cellular response to inflammatory cytokines, environmental stress and pathogenic infection, However, compared with mammals and insects, the potential function of p38 MAPK from Chinese shrimp (Fenneropenaeus chinensis) in response to viruses and bacterial infection is limited. In the study, the immune responses of four genes MKK3, MKK4, p38 and ATF-2 from F. chinensis were investigated in defending against white spot syndrome virus (WSSV), Vibrio anguillarum (V...
November 16, 2017: Gene
https://www.readbyqxmd.com/read/29155083/tbvps15-is-required-for-vesicular-transport-and-cytokinesis-in-trypanosoma-brucei
#10
Alejandra C Schoijet, Kildare Miranda, Tamara Sternlieb, Nadia M Barrera, Wendell Girard-Dias, Wanderley de Souza, Guillermo D Alonso
The class III phosphatidylinositol 3-kinase (PI3K) Vps34 is an important regulator of key cellular functions, including cell growth, survival, intracellular trafficking, autophagy and nutrient sensing. In yeast, Vps34 is associated with the putative serine/threonine protein kinase Vps15, however, its role in signaling has not been deeply evaluated. Here, we have identified the Vps15 orthologue in Trypanosoma brucei, named TbVps15. Knockdown of TbVps15 expression by interference RNA resulted in inhibition of cell growth and blockage of cytokinesis...
November 16, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/29150005/crispr-libraries-and-screening
#11
John T Poirier
CRISPR-Cas9 technology has revolutionized large-scale functional genomic screening in mammalian cell-culture systems. Due in part to optimized lentiviral delivery vectors; it is now possible to perform CRISPR-Cas9 screens in animals in order to study biological processes in the context of a whole organism and within more physiologically relevant environment. This chapter focuses primarily on mouse models of human cancers; viral vectors used for simultaneous tumor initiation and genome editing and sgRNA library design considerations...
2017: Progress in Molecular Biology and Translational Science
https://www.readbyqxmd.com/read/29149987/inhibition-of-porcine-endogenous-retrovirus-by-multi-targeting-micro-rna-against-long-terminal-region
#12
H-C Chung, V-G Nguyen, W-T Oh, T-M-L Huynh, H-J Moon, J-H Lee, H-K Kim, S-J Park, B-K Park
BACKGROUND: There might be much benefit in xenotransplantation, however, the risk of infections across species barriers remains, especially porcine endogenous retrovirus (PERV). To date, many attempts have been made to knock down active PERVs by inhibitory RNA (RNAi) and micro RNA (miRNA), which target different genes of PERV. There are a few studies that have explored whether targeting promoter regions of PERV could exert an inhibition effect. METHODS: miRNAs were automatically selected based on an online program BLOCK-iT RNAi Designer...
November 2017: Transplantation Proceedings
https://www.readbyqxmd.com/read/29149417/spl13-regulates-shoot-branching-and-flowering-time-in-medicago-sativa
#13
Ruimin Gao, Margaret Y Gruber, Lisa Amyot, Abdelali Hannoufa
Our results show SPL13 plays a crucial role in regulating vegetative and reproductive development in Medicago sativa L. (alfalfa), and that MYB112 is targeted and downregulated by SPL13 in alfalfa. We previously showed that transgenic Medicago sativa (alfalfa) plants overexpressing microRNA156 (miR156) show a bushy phenotype, reduced internodal length, delayed flowering time, and enhanced biomass yield. In alfalfa, transcripts of seven SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors, including SPL13, are targeted for cleavage by miR156...
November 17, 2017: Plant Molecular Biology
https://www.readbyqxmd.com/read/29148850/atad2-in-cancer-a-pharmacologically-challenging-but-tractable-target
#14
Muzammal Hussain, Yang Zhou, Yu Song, H M Adnan Hameed, Hao Jiang, Yaoquan Tu, Jiancun Zhang
ATAD2 protein is an emerging oncogene that has strongly been linked to the etiology of multiple advanced human cancers. Therapeutically, despite the fact that genetic suppression/knockdown studies have validated it as a compelling drug target for future therapeutic development, recent druggability assessment data suggest that direct targeting of ATAD2's bromodomain (BRD) may be a very challenging task. ATAD2's BRD has been predicted as a "difficult to drug" or "least druggable" target due to the concern that its binding pocket, and the areas around it, seem to be unfeasible for ligand binding...
November 17, 2017: Expert Opinion on Therapeutic Targets
https://www.readbyqxmd.com/read/29145490/rnai-screen-reveals-a-role-of-sphk2-in-dengue-virus-mediated-apoptosis-in-hepatic-cell-lines
#15
Atthapan Morchang, Regina Ching Hua Lee, Pa-Thai Yenchitsomanus, Gopinathan Pillai Sreekanth, Sansanee Noisakran, Justin Jang Hann Chu, Thawornchai Limjindaporn
Hepatic dysfunction is a feature of dengue virus (DENV) infection. Hepatic biopsy specimens obtained from fatal cases of DENV infection show apoptosis, which relates to the pathogenesis of DENV infection. However, how DENV induced liver injury is not fully understood. In this study, we aim to identify the factors that influence cell death by employing an apoptosis-related siRNA library screening. Our results show the effect of 558 gene silencing on caspase 3-mediated apoptosis in DENV-infected Huh7 cells. The majority of genes that contributed to apoptosis were the apoptosis-related kinase enzymes...
2017: PloS One
https://www.readbyqxmd.com/read/29144989/cysteine-dioxygenase-1-mediates-erastin-induced-ferroptosis-in-human-gastric-cancer-cells
#16
Shihui Hao, Jiang Yu, Wanming He, Qiong Huang, Yang Zhao, Bishan Liang, Shuyi Zhang, Zhaowei Wen, Shumin Dong, Jinjun Rao, Wangjun Liao, Min Shi
BACKGROUND: Ferroptosis is a recently discovered form of iron-dependent nonapoptotic cell death. It is characterized by loss of the activity of the lipid repair enzyme, glutathione peroxidase 4 (GPX4), and accumulation of lethal reactive lipid oxygen species. However, we still know relatively little about ferroptosis and its molecular mechanism in gastric cancer (GC) cells. Here, we demonstrate that erastin, a classic inducer of ferroptosis, induces this form of cell death in GC cells and that cysteine dioxygenase 1 (CDO1) plays an important role in this process...
November 13, 2017: Neoplasia: An International Journal for Oncology Research
https://www.readbyqxmd.com/read/29144122/the-ratio-of-polycation-and-serum-is-a-crucial-index-determining-the-rnai-efficiency-of-polyplex
#17
Weiqi Zhang, Xianghui Meng, Huike Liu, Lifei Xie, Jian Liu, Haiyan Xu
We report that the mass ratio of the polycation to serum in the medium determines the RNAi efficiency in vitro by using spermine-modified pullulan (Ps) and spermine-modified dextran (Ds) as polycation models. The high ratio of Ps to serum protein (Ps/Pr) mediated the formation of larger polyplex, which led to the promoted cellular uptake, enhanced lysosomal escape and elevated RNAi efficiency. In addition, the supplementary of free Ps also enhanced siRNA transfection due to the elevation of Ps/Pr. Similar results were obtained with Ds...
November 16, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29142303/fus-toxicity-is-rescued-by-the-modulation-of-lncrna-hsr%C3%AF-expression-in-drosophila-melanogaster
#18
Luca Lo Piccolo, Salinee Jantrapirom, Yoshitaka Nagai, Masamitsu Yamaguchi
FUS is an aggregation-prone hnRNP involved in transcriptional and post-transcriptional regulation that aberrantly forms immunoreactive inclusion bodies in a range of neurological diseases classified as FUS-proteinopathies. Although FUS has been extensively examined, the underlying molecular mechanisms of these diseases have not yet been elucidated in detail. We previously reported that RNAi of the lncRNA hsrω altered the expression and sub-cellular localization of Drosophila FUS in the central nervous system of the fly...
November 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29141233/mirnome-profiling-of-purified-endoderm-and-mesoderm-differentiated-from-hescs-reveals-functions-of-mir-483-3p-and-mir-1263-for-cell-fate-decisions
#19
Daichi Ishikawa, Ulf Diekmann, Jan Fiedler, Annette Just, Thomas Thum, Sigurd Lenzen, Ortwin Naujok
Pluripotent stem cells hold great promise for regenerative medicine since they can differentiate into all somatic cells. MicroRNAs (miRNAs) could be important for the regulation of these cell-fate decisions. Profiling of miRNAs revealed 19 differentially expressed miRNAs in the endoderm and 29 in the mesoderm when analyzing FACS-purified cells derived from human embryonic stem cells. The mesodermal-enriched miR-483-3p was identified as an important regulator for the generation of mesodermal PDGFRA(+) paraxial cells...
November 14, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/29139179/involvement-of-lemrp-an-atp-binding-cassette-transporter-in-shikonin-transport-and-biosynthesis-in-lithospermum-erythrorhizon
#20
Yu Zhu, Shu-Juan Chu, Yue-Lin Luo, Jiang-Yan Fu, Cheng-Yi Tang, Gui-Hua Lu, Yan-Jun Pang, Xiao-Ming Wang, Rong-Wu Yang, Jin-Liang Qi, Yong-Hua Yang
·Shikonin and its derivatives are important medicinal secondary metabolites accumulating in the roots of Lithospermum erythrorhizon. Although some membrane proteins have been identified as transporters of secondary metabolites, the mechanisms underlying shikonin transport and accumulation in L. erythrorhizon cells still remain largely unknown. ·In this study, we isolated a cDNA encoding LeMRP, an ATP-binding cassette transporter from L. erythrorhizon and further investigated its functions in the transport and biosynthesis of shikonin with the yeast transformation and transgenic hairy root method, respectively...
November 15, 2017: Plant Biology
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