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Ru-Yi Yu, Lei Xing, Peng-Fei Cui, Jian-Bin Qiao, Yu-Jing He, Xin Chang, Tian-Jiao Zhou, Qing-Ri Jin, Hu-Lin Jiang, Yanyu Xiao
Finding a cure for breast cancer currently remains a medical challenge in due to the failure of common treatment methods to inhibit invasion and metastasis of cancer cells, which eventually leads to recurrence of breast cancer. Many secreted proteins are overexpressed and play crucial roles in tumorigenesis and development. The Golgi apparatus is a key protein processing and secretion factory in which metastasis-associated proteins are modified, transported and secreted; thus, regulating the Golgi apparatus of tumor cells is a viable strategy to inhibit tumor metastasis...
June 20, 2018: Biomaterials Science
Shao-Ming Shen, Yan Ji, Cheng Zhang, Shuang-Shu Dong, Shuo Yang, Zhong Xiong, Meng-Kai Ge, Yun Yu, Li Xia, Meng Guo, Jin-Ke Cheng, Jun-Ling Liu, Jian-Xiu Yu, Guo-Qiang Chen
Dysregulation of pre-mRNA alternative splicing (AS) is closely associated with cancers. However, the relationships between the AS and classic oncogenes/tumor suppressors are largely unknown. Here we show that the deletion of tumor suppressor PTEN alters pre-mRNA splicing in a phosphatase-independent manner, and identify 262 PTEN-regulated AS events in 293T cells by RNA sequencing, which are associated with significant worse outcome of cancer patients. Based on these findings, we report that nuclear PTEN interacts with the splicing machinery, spliceosome, to regulate its assembly and pre-mRNA splicing...
June 19, 2018: Nature Communications
Nora E Gray, Jonathan A Zweig, Maya Caruso, Marjoen D Martin, Jennifer Y Zhu, Joseph F Quinn, Amala Soumyanath
INTRODUCTION: Centella asiatica is a plant used for centuries to enhance memory. We have previously shown that a water extract of Centella asiatica (CAW) attenuates age-related spatial memory deficits in mice and improves neuronal health. Yet the effect of CAW on other cognitive domains remains unexplored as does its mechanism of improving age-related cognitive impairment. This study investigates the effects of CAW on a variety of cognitive tasks as well as on synaptic density and mitochondrial and antioxidant pathways...
June 19, 2018: Brain and Behavior
Qingtian Li, Fan Lei, Yi Tang, Jenny Szu-Chin Pan, Qiang Tong, Yuxiang Sun, David Sheikh-Hamad
Mitochondrial intracrines are extracellular signaling proteins, targeted to the mitochondria. The pathway for mitochondrial targeting of mitochondrial intracrines and actions in the mitochondria remains unknown. Megalin/LRP2 mediates the uptake of vitamins and proteins, and is critical for clearance of amyloid-β protein from the brain. Megalin mutations underlie the pathogenesis of Donnai-Barrow and Lowe syndromes, characterized by brain defects and kidney dysfunction; megalin was not previously known to reside in the mitochondria...
June 9, 2018: Cellular and Molecular Life Sciences: CMLS
Junpei Takagi, Tomohiro Uemura
Eukaryotic cells comprise various organelles surrounded by the membrane. Each organelle is characterized by unique proteins and lipids and has its own specific functions. Single membrane-bounded organelles, including the Golgi apparatus, endosomes, and vacuoles are connected by membrane trafficking. Identifying the organelle localization of a protein of interest is essential for determining the proteins physiological functions. Here, we describe methods for determining protein subcellular localization using the inhibitors brefeldin A and wortmannin in Arabidopsis thaliana...
2018: Methods in Molecular Biology
Dominik P Buser, Kai D Schleicher, Cristina Prescianotto-Baschong, Martin Spiess
Retrograde transport of membranes and proteins from the cell surface to the Golgi and beyond is essential to maintain homeostasis, compartment identity, and physiological functions. To study retrograde traffic biochemically, by live-cell imaging or by electron microscopy, we engineered functionalized anti-GFP nanobodies (camelid VHH antibody domains) to be bacterially expressed and purified. Tyrosine sulfation consensus sequences were fused to the nanobody for biochemical detection of trans -Golgi arrival, fluorophores for fluorescence microscopy and live imaging, and APEX2 (ascorbate peroxidase 2) for electron microscopy and compartment ablation...
June 18, 2018: Proceedings of the National Academy of Sciences of the United States of America
Bo Am Seo, Taesup Cho, Daniel Z Lee, Joong-Jae Lee, Boyoung Lee, Seong-Wook Kim, Hee-Sup Shin, Myoung-Goo Kang
Mutations in the human LARGE gene result in severe intellectual disability and muscular dystrophy. How LARGE mutation leads to intellectual disability, however, is unclear. In our proteomic study, LARGE was found to be a component of the AMPA-type glutamate receptor (AMPA-R) protein complex, a main player for learning and memory in the brain. Here, our functional study of LARGE showed that LARGE at the Golgi apparatus (Golgi) negatively controlled AMPA-R trafficking from the Golgi to the plasma membrane, leading to down-regulated surface and synaptic AMPA-R targeting...
June 18, 2018: Proceedings of the National Academy of Sciences of the United States of America
Antonietta Rosella Farina, Lucia Cappabianca, Pierdomenico Ruggeri, Luciana Gneo, Cristina Pellegrini, Maria-Concetta Fargnoli, Andrew Reay Mackay
Oncogenes derived from the neurotrophin receptor tropomyosin-related kinase TrkA act as drivers in sub-populations of a wide-range of human cancers. This, combined with a recent report that both adult and childhood cancers driven by novel oncogenic TrkA chimeric-fusions exhibit profound, long-lived therapeutic responses to the Trk inhibitor Larotrectinib, highlights the need to improve clinical detection of TrkA oncogene-driven cancers in order to maximise this novel therapeutic potential. Cancers potentially driven by TrkA oncogenes include a proportion of paediatric neuroblastomas (NBs) that express the alternative TrkA splice variant TrkAIII, which exhibits exon 6, 7 and 9 skipping and oncogenic-activity that depends upon deletion of the extracellular D4 Ig-like domain...
June 18, 2018: Journal of Experimental & Clinical Cancer Research: CR
Hongying Liu, Xieqi Wang, Bing Feng, Lipeng Tang, Weiping Li, Xirun Zheng, Ying Liu, Yan Peng, Guangjuan Zheng, Qinglian He
BACKGROUND: Hepatocellular carcinoma (HCC) is the sixth most common cancer and the second leading cause of cancer-related deaths worldwide. Despite new technologies in diagnosis and treatment, the incidence and mortality of HCC continue rising. And its pathogenesis is still unclear. As a highly conserved protein of the Golgi apparatus, Golgi phosphoprotein 3 (GOLPH3) has been shown to be involved in tumorigenesis of HCC. This study aimed to explore the exact oncogenic mechanism of GOLPH3 and provide a novel diagnose biomarker and therapeutic strategy for patients with HCC...
June 18, 2018: BMC Cancer
Hyun Jin Cho, Inhee Mook-Jung
The conventional secretory pathway is indispensable for eukaryotic cells. Newly synthesized membrane and secretory proteins are released from the endoplasmic reticulum (ER) through ER-derived vesicles to their appropriate destination. Vesicle formation is important for steady protein trafficking. O-GlcNAcylation ( O-GlcNAc) is a unique protein glycosylation signature, whose dynamic regulation by O-GlcNAc transferase and O-GlcNAcase occurs exclusively for nuclear and cytoplasmic proteins. Because of this locally limited property, the role of O-GlcNAc in the conventional protein secretory pathway is unknown...
April 17, 2018: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Jolanta Zegarlińska, Magda Piaścik, Aleksander F Sikorski, Aleksander Czogalla
Phosphatidic acid (PA) is the simplest glycerophospholipid naturally occurring in living organisms, and even though its content among other cellular lipids is minor, it is drawing more and more attention due to its multiple biological functions. PA is a precursor for other phospholipids, acts as a lipid second messenger and, due to its structural properties, is also a modulator of membrane shape. Although much is known about interaction of PA with its effectors, the molecular mechanisms remain unresolved to a large degree...
2018: Acta Biochimica Polonica
Eric J M Blondeel, Marc G Aucoin
Glycosylation is a critical quality attribute (CQA) of many therapeutic proteins, particularly monoclonal antibodies (mAbs), and is a major consideration in the approval of biosimilar biologics due to its effects to therapeutic efficacy. Glycosylation generates a distribution of glycoforms, resulting in glycoproteins with inherent molecule-to-molecule heterogeneity, capable of activating (or failing to activate) different effector functions of the immune system. Glycoforms can be affected by the supplementation of nucleotide-sugar precursors, and related components, to culture growth medium, affecting the metabolism of glycosylation...
June 15, 2018: Biotechnology Advances
Aqeela Zahra, Jinxiang Jiang, Yunan Chen, Cheng Long, Li Yang
Prenatal exposure to citalopram (CTM), an antidepressant drug, has been associated with altered behavior, including autism-like symptoms in both human and rodent offspring. However, the neurological basis underlying these abnormal behaviors is not well understood. Here, we examined behavioral, morphological, and biochemical alterations in the male and female offspring of C57BL/6 mouse mothers that had been exposed to CTM during the last trimester of gestation. We observed abnormal behavior such as anxiety, altered locomotion and disordered social interactions in 2-5 month old offspring with prenatal CTM exposure...
June 15, 2018: Experimental Neurology
Tatsuki Kikegawa, Takuya Yamaguchi, Ryohei Nambu, Kenji Etchuya, Masami Ikeda, Yuri Mukai
Despite studies of the mechanism underlying the intracellular localization of membrane proteins, the specific mechanisms by which each membrane protein localizes to the endoplasmic reticulum, Golgi apparatus, and plasma membrane in the secretory pathway are unclear. In this study, a discriminant analysis of endoplasmic reticulum, Golgi apparatus and plasma membrane-localized type II membrane proteins was performed using a position-specific scoring matrix derived from the amino acid propensity of the sequences around signal-anchors...
June 18, 2018: Bioscience, Biotechnology, and Biochemistry
Xin Yong, Wenfeng Hu, Xue Zhou, Jing Wang, Ezra Burstein, Da Jia
The sorting nexin (SNX) family proteins play an essential role in vesicular transport, cell signaling, and membrane remodeling. The SNX members SNX1/2 and SNX5/6 form dimers, and mediate endosome-to-trans Golgi network (TGN) transport through coordinating cargo selection and membrane remodeling. It is well-known how a SNX-BAR protein forms a homodimer; however, it is less clear how a heterodimer is formed. Here a detailed expression and purification protocol of the SNX1/SNX6 complex, from both worm and human, is described...
June 14, 2018: Protein Expression and Purification
Markus R M Fiedler, Lars Barthel, Christin Kubisch, Corrado Nai, Vera Meyer
BACKGROUND: The lifestyle of filamentous fungi depends on the secretion of hydrolytic enzymes into the surrounding medium, which degrade polymeric substances into monomers that are then taken up to sustain metabolism. This feature has been exploited in biotechnology to establish platform strains with high secretory capacity including Aspergillus niger. The accepted paradigm is that proteins become mainly secreted at the tips of fungal hyphae. However, it is still a matter of debate if the amount of growing hyphal tips in filamentous fungi correlates with an increase in secretion, with previous studies showing either a positive or no correlation...
June 16, 2018: Microbial Cell Factories
Ronaldo Roberto Tait Caleffe, Stefany Rodrigues de Oliveira, Adriana Aparecida Sinópolis Gigliolli, Maria Claudia Colla Ruvolo-Takasusuki, Helio Conte
Larval therapy (LT) comprises the application of sterile Calliphoridae larvae for wound debridement, disinfection, and healing in humans and animals. Larval digestion plays a key role in LT, where the salivary glands and gut produce and secrete proteolytic and antimicrobial substances. The objective of this work was to bioprospect the salivary glands of Chrysomya megacephala (Fabricius, 1794) larvae, using ultrastructural, morphological, and histological observations, and the total protein electrophoretic profile...
June 8, 2018: Micron: the International Research and Review Journal for Microscopy
Romina Canzoneri, Martín Enrique Rabassa, Agustina Gurruchaga, Valeria Ferretti, Sabina Palma, Marina Isla-Larrain, María Virginia Croce, Ezequiel Lacunza, Martín Carlos Abba
RHBDD2 is an intramembrane pseudoprotease member of the Rhomboid superfamily. Our previous studies in breast and colorectal cancer indicate an association between RHBDD2 overexpression and advanced tumor stages. Two alternative transcriptional variants have been described for RHBDD2, which would be encoding for different RHBDD2 protein isoforms. The expression of these RHBDD2 variants/isoforms and its association with breast cancer was the focus of this study. First, expression of RHBDD2 splicing variants was evaluated in normal and breast tumor samples...
June 14, 2018: Oncology Reports
Masahiro Kumeta, Yulia Panina, Hiroya Yamazaki, Kunio Takeyasu, Shige H Yoshimura
The primary cilium functions as an "antenna" for cell signaling, studded with characteristic transmembrane receptors and soluble protein factors, raised above the cell surface. In contrast to the transmembrane proteins, targeting mechanisms of nontransmembrane ciliary proteins are poorly understood. We focused on a pathogenic mutation that abolishes ciliary localization of retinitis pigmentosa 2 protein and revealed a dual acylation-dependent ciliary targeting pathway. Short N-terminal sequences which contain myristoylation and palmitoylation sites are sufficient to target a marker protein into the cilium in a palmitoylation-dependent manner...
June 13, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Midori Ishii, Vladimir V Lupashin, Akihiko Nakano
The Golgi apparatus is a central station for protein trafficking in eukaryotic cells. A widely accepted model of protein transport within the Golgi apparatus is cisternal maturation. Each cisterna has specific resident proteins, which are thought to be maintained by COPI-mediated transport. However, the mechanisms underlying specific sorting of these Golgi-resident proteins remain elusive. To obtain a clue to understand the selective sorting of vesicles between the Golgi cisterenae, we investigated the molecular arrangements of the conserved oligomeric Golgi (COG) subunits in yeast cells...
June 14, 2018: Cell Structure and Function
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