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https://www.readbyqxmd.com/read/29346269/identification-of-secreted-proteins-involved-in-nonspecific-dsrna-mediated-lutzomyia-longipalpis-ll5-cell-antiviral-response
#1
Andrea Martins-da-Silva, Erich Loza Telleria, Michel Batista, Fabricio Klerynton Marchini, Yara Maria Traub-Csekö, Antonio Jorge Tempone
Hematophagous insects transmit infectious diseases. Sand flies are vectors of leishmaniasis, but can also transmit viruses. We have been studying immune responses of Lutzomyia longipalpis, the main vector of visceral leishmaniasis in the Americas. We identified a non-specific antiviral response in L. longipalpis LL5 embryonic cells when treated with non-specific double-stranded RNAs (dsRNAs). This response is reminiscent of interferon response in mammals. We are investigating putative effectors for this antiviral response...
January 18, 2018: Viruses
https://www.readbyqxmd.com/read/29316283/split-cas9-not-hairs-advancing-the-therapeutic-index-of-crispr-technology
#2
REVIEW
Carolin Schmelas, Dirk Grimm
The discovery that the bacterial CRISPR/Cas9 system can be translated into mammalian cells continues to have an unprecedented impact on the biomedical research community, as it largely facilitates efforts to experimentally interrogate or therapeutically modify the cellular genome. In particular, CRISPR promises the ability to correct disease-associated genetic defects, or to target and destroy invading foreign DNA, in a simple, efficient and selective manner directly in affected human cells or tissues. Here, we highlight a set of exciting new strategies that aim at further increasing the therapeutic index of CRISPR technologies, by reducing the size of Cas9 expression cassettes and thus enhancing their compatibility with viral gene delivery vectors...
January 5, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29246765/fkbp12-immunopositive-inclusions-in-patients-with-%C3%AE-synucleinopathies
#3
Yasuyuki Honjo, Takashi Ayaki, Tomohisa Horibe, Hidefumi Ito, Ryosuke Takahashi, Koji Kawakami
α-Synuclein (α-SYN), a presynaptic protein with the tendency to aggregate, is linked to α-synucleinopathies such as Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). α-SYN is the main component of round intracytoplasmic inclusions called Lewy bodies (LBs), which are the hallmark of PD and DLB. In addition, accumulation of amyloid-β and neurofibrillary tangles as in the pathology of Alzheimer's disease has been found in the DLB brain. Glial cytoplasmic inclusions are an MSA-specific type of inclusion found in oligodendrocytes and mainly comprise α-SYN...
December 12, 2017: Brain Research
https://www.readbyqxmd.com/read/29206196/the-fkbp51-glucocorticoid-receptor-co-chaperone-regulation-function-and-implications-in-health-and-disease
#4
REVIEW
Gabriel R Fries, Nils C Gassen, Theo Rein
Among the chaperones and co-chaperones regulating the glucocorticoid receptor (GR), FK506 binding protein (FKBP) 51 is the most intensely investigated across different disciplines. This review provides an update on the role of the different co-chaperones of Hsp70 and Hsp90 in the regulation of GR function. The development leading to the focus on FKBP51 is outlined. Further, a survey of the vast literature on the mechanism and function of FKBP51 is provided. This includes its structure and biochemical function, its regulation on different levels-transcription, post-transcription, and post-translation-and its function in signaling pathways...
December 5, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29194511/prolyl-isomerization-of-the-cenp-a-n-terminus-regulates-centromeric-integrity-in-fission-yeast
#5
Hwei Ling Tan, Kim Kiat Lim, Qiaoyun Yang, Jing-Song Fan, Ahmed Mahmoud Mohammed Sayed, Liy Sim Low, Bingbing Ren, Teck Kwang Lim, Qingsong Lin, Yu-Keung Mok, Yih-Cherng Liou, Ee Sin Chen
Centromeric identity and chromosome segregation are determined by the precise centromeric targeting of CENP-A, the centromere-specific histone H3 variant. The significance of the amino-terminal domain (NTD) of CENP-A in this process remains unclear. Here, we assessed the functional significance of each residue within the NTD of CENP-A from Schizosaccharomyces pombe (SpCENP-A) and identified a proline-rich 'GRANT' (Genomic stability-Regulating site within CENP-A N-Terminus) motif that is important for CENP-A function...
November 29, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29155566/structure-based-insights-into-dynamics-and-function-of-two-domain-slpa-from-escherichia-coli
#6
Anne-Juliane Geitner, Ulrich Weininger, Hauke Paulsen, Jochen Balbach, Michael Kovermann
SlpA (SlyD like protein A) comprises two domains, a FK506 binding domain (FKBP fold) of moderate prolyl cis/trans isomerase activity and an inserted in flap (IF) domain that hosts its chaperone activity. Here we present the NMR solution structure of apo E. coli SlpA determined by NMR which mirrors the structural properties seen for various SlyD homologs. Crucial structural differences in side-chain orientation arise for F37 which points directly into the hydrophobic core of the active site. It forms a prominent aromatic stacking with F15, one of the key residues for PPIase activity, thus giving a possible explanation for the inherently low PPIase activity of SlpA...
November 20, 2017: Biochemistry
https://www.readbyqxmd.com/read/29148176/differential-proteomic-analysis-reveals-protein-networks-and-pathways-that-may-contribute-to-helicobacter-pylori-fkbp-type-ppiase-associated-gastric-diseases
#7
Yanmei Zhu, Yuehua Gong, Aodi Li, Moye Chen, Dan Kang, Jun Liu, Yuan Yuan
PURPOSE: Though Helicobacter pylori (H. pylori) has been classified as class I carcinogen, key virulence factor generated by H. pylori that causes gastric cancer remains to be fully determined. Recently, we identified a gastric cancer-associated H. pylori gene, peptidylprolyl isomerase-FK506 binding protein (PPIase-FKBP), and showed that PPIase-FKBP was capable of inducing oncogenic transformation of gastric epithelial cells. But its mechanism was unclear. EXPERIMENTAL DESIGN: We carried out a comparative proteomic analysis of human gastric epithelial cells that either express PPIase-FKBP or green fluorescent protein (GFP) using two-dimensional (2-D) gel electrophoresis and then matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS/MS)...
November 16, 2017: Proteomics. Clinical Applications
https://www.readbyqxmd.com/read/29115621/effects-of-shield1-on-the-viral-replication-of-varicella%C3%A2-zoster-virus-containing-fkbp%C3%A2-tagged-orf4-and-48
#8
Shuying Li, Zhanjun Liu, Ji Li, Aihua Liu, Lihua Zhu, Kui Yu, Ke Zhang
The present study aimed to explore the effects of a stabilizing ligand, Shield‑1, on the replication of recombinant varicella‑zoster virus (VZV) containing FK506 binding protein (FKPB) tags in essential open reading frames (ORF) 4 and 48. A specific galactokinase (galK) selection method was conducted, following the addition of galK labels to VZV ORF4 and 48, using a SW102 VZV bacterial artificial chromosome (BAC) system. Subsequently, recombinant VZV containing FKPB tags in ORF4 and 48 was constructed by counterselection and homologous recombination...
November 6, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29109782/berunda-polypeptides-multi-headed-fusion-proteins-promote-subcutaneous-administration-of-rapamycin-to-breast-cancer-in-vivo
#9
Jugal P Dhandhukia, Zhe Li, Santosh Peddi, Shruti Kakan, Arjun Mehta, David Tyrpak, Jordan Despanie, J Andrew MacKay
Recombinant Elastin-Like Polypeptides (ELPs) serve as attractive scaffolds for nanoformulations because they can be charge-neutral, water soluble, high molecular weight, monodisperse, biodegradable, and decorated with functional proteins. We recently reported that fusion of the FK-506 binding protein 12 (FKBP) to an ELP nanoparticle (FSI) reduces rapamycin (Rapa) toxicity and enables intravenous (IV) therapy in both a xenograft breast cancer model and a murine autoimmune disease model. Rapa has poor solubility, which leads to variable oral bioavailability or drug precipitation following parenteral administration...
2017: Theranostics
https://www.readbyqxmd.com/read/29073761/targeted-gene-repression-using-novel-bifunctional-molecules-to-harness-endogenous-histone-deacetylation-activity
#10
Kyle V Butler, Anna M Chiarella, Jian Jin, Nathaniel A Hathaway
Epigenome editing is a powerful method for life science research and could give rise to new therapies for diseases initiated or maintained by epigenetic dysregulation, including several types of cancers and autoimmune disorders. In addition, much is still unknown about the mechanisms by which histone-modifying proteins work in concert to properly regulate gene expression. To investigate and manipulate complex epigenetic interactions in live cells, we have developed a small molecule platform for specifically inducing gene repression and histone deacetylation at a reporter gene...
November 7, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/29042512/in-vitro-reconstitution-of-t-cell-receptor-mediated-segregation-of-the-cd45-phosphatase
#11
Catherine B Carbone, Nadja Kern, Ricardo A Fernandes, Enfu Hui, Xiaolei Su, K Christopher Garcia, Ronald D Vale
T cell signaling initiates upon the binding of peptide-loaded MHC (pMHC) on an antigen-presenting cell to the T cell receptor (TCR) on a T cell. TCR phosphorylation in response to pMHC binding is accompanied by segregation of the transmembrane phosphatase CD45 away from TCR-pMHC complexes. The kinetic segregation hypothesis proposes that CD45 exclusion shifts the local kinase-phosphatase balance to favor TCR phosphorylation. Spatial partitioning may arise from the size difference between the large CD45 extracellular domain and the smaller TCR-pMHC complex, although parsing potential contributions of extracellular protein size, actin activity, and lipid domains is difficult in living cells...
October 31, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29036638/the-basic-tilted-helix-bundle-domain-of-the-prolyl-isomerase-fkbp25-is-a-novel-double-stranded-rna-binding-module
#12
David Dilworth, Santosh K Upadhyay, Pierre Bonnafous, Amiirah Bibi Edoo, Sarah Bourbigot, Francy Pesek-Jardim, Geoff Gudavicius, Jason J Serpa, Evgeniy V Petrotchenko, Christoph H Borchers, Christopher J Nelson, Cameron D Mackereth
Prolyl isomerases are defined by a catalytic domain that facilitates the cis-trans interconversion of proline residues. In most cases, additional domains in these enzymes add important biological function, including recruitment to a set of protein substrates. Here, we report that the N-terminal basic tilted helix bundle (BTHB) domain of the human prolyl isomerase FKBP25 confers specific binding to double-stranded RNA (dsRNA). This binding is selective over DNA as well as single-stranded oligonucleotides. We find that FKBP25 RNA-association is required for its nucleolar localization and for the vast majority of its protein interactions, including those with 60S pre-ribosome and early ribosome biogenesis factors...
November 16, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29032102/regulation-of-ca-2-signaling-by-acute-hypoxia-and-acidosis-in-rat-neonatal-cardiomyocytes
#13
José-Carlos Fernández-Morales, Martin Morad
Ischemic heart disease is an arrhythmogenic condition, accompanied by hypoxia, acidosis, and impaired Ca(2+) signaling. Here we report on effects of acute hypoxia and acidification in rat neonatal cardiomyocytes cultures. RESULTS: Two populations of neonatal cardiomyocyte were identified based on inactivation kinetics of L-type ICa: rapidly-inactivating ICa (τ~20ms) myocytes (prevalent in 3-4-day cultures), and slow-inactivating ICa (τ≥40ms) myocytes (dominant in 7-day cultures). Acute hypoxia (pO2<5mmHg for 50-100s) suppressed ICa reversibly in both cell-types to different extent and with different kinetics...
October 12, 2017: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/29024217/use-of-the-target-of-rapamycin-fatc-domain-as-a-general-membrane-anchor-the-fkbp-12-like-domain-of-fkbp38-as-a-case-study
#14
Maristella De Cicco, Lech-G Milroy, Sonja A Dames
Increased efforts have been undertaken to better understand the formation of signaling complexes at cellular membranes. Since the preparation of proteins containing a transmembrane domain or a prenylation motif is generally challenging an alternative membrane anchoring unit that is easy to attach, water-soluble and binds to different membrane mimetics would find broad application. The 33-residue long FATC domain of yeast TOR1 (y1fatc) fulfills these criteria and binds to neutral and negatively charged micelles, bicelles and liposomes...
October 12, 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28982713/publisher-s-note-association-of-fk506-binding-proteins-with-ryr-channels-effect-of-clic2-binding-on-sub-conductance-opening-and-fkbp-binding-j-cell-sci-doi-10-1242-jcs-204461
#15
Spencer J Richardson, Gregory A Steele, Esther M Gallant, Alexander Lam, Charles E Schwartz, Philip G Board, Marco G Casarotto, Nicole A Beard, Angela F Dulhunty
No abstract text is available yet for this article.
October 5, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28973376/the-integrity-and-organization-of-the-human-aipl1-functional-domains-is-critical-for-its-role-as-a-hsp90-dependent-co-chaperone-for-rod-pde6
#16
Almudena Sacristan-Reviriego, James Bellingham, Chrisostomos Prodromou, Neruban Kumaran, James Bainbridge, Michel Michaelides, Jacqueline van der Spuy
Biallelic mutations in the photoreceptor-expressed aryl hydrocarbon receptor interacting protein-like 1 (AIPL1) are associated with autosomal recessive Leber congenital amaurosis (LCA), the most severe form of inherited retinopathy in early childhood. AIPL1 functions as a photoreceptor-specific co-chaperone that interacts with the molecular chaperone HSP90 to facilitate the stable assembly of the retinal cyclic GMP (cGMP) phosphodiesterase (PDE6) holoenzyme. In this study, we characterized the functional deficits of AIPL1 variations, some of which induce aberrant pre-mRNA AIPL1 splicing leading to the production of alternative AIPL1 isoforms...
November 15, 2017: Human Molecular Genetics
https://www.readbyqxmd.com/read/28965803/expression-purification-and-characterization-of-a-catalytic-domain-of-human-protein-tyrosine-phosphatase-non-receptor-12-ptpn12-in-escherichia-coli-with-fkbp-type-ppiase-as-a-chaperon
#17
Yuan Sui, Xingye Fu, Yuchen Wang, Weiyan Hu, Tong Zhang, Wanyao Liu, Liyan Jiang, Shu Xing, Xueqi Fu, Xuesong Xu
Protein tyrosine phosphatase non-receptor type 12 (PTPN12), also known as PTP-PEST, was broadly expressed in hemopoietic cells. Recent research has shown that this enzyme is involved in tumorigenesis, as well as in tumor progression and transfer, as it can suppress multiple oncogenic tyrosine kinases. However, the difficulty of soluble expression of PTP-PEST in prokaryotic cells has resulted in great limitations in investigating its structure and functions. In this study, we successfully carried out soluble expression of the catalytic domain of PTP-PEST (ΔPTP-PEST) in Escherichia coli and performed an enzymatic characterization and kinetics...
September 28, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28955036/optimizing-the-fragment-complementation-of-apex2-for-detection-of-specific-protein-protein-interactions-in-live-cells
#18
Miaomiao Xue, Junjie Hou, Linlin Wang, Dongwan Cheng, Jingze Lu, Li Zheng, Tao Xu
Dynamic protein-protein interactions (PPIs) play crucial roles in cell physiological processes. The protein-fragment complementation (PFC) assay has been developed as a powerful approach for the detection of PPIs, but its potential for identifying protein interacting regions is not optimized. Recently, an ascorbate peroxidase (APEX2)-based proximity-tagging method combined with mass spectrometry was developed to identify potential protein interactions in live cells. In this study, we tested whether APEX2 could be employed for PFC...
September 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28937754/bifunctional-elastin-like-polypeptide-nanoparticles-bind-rapamycin-and-integrins-and-suppress-tumor-growth-in-vivo
#19
Jugal P Dhandhukia, Pu Shi, Santosh Peddi, Zhe Li, Suhaas Aluri, Yaping Ju, Dab Brill, Wan Wang, Siti M Janib, Yi-An Lin, Shuanglong Liu, Honggang Cui, J Andrew MacKay
Recombinant protein-polymer scaffolds such as elastin-like polypeptides (ELPs) offer drug-delivery opportunities including biocompatibility, monodispersity, and multifunctionality. We recently reported that the fusion of FK-506 binding protein 12 (FKBP) to an ELP nanoparticle (FSI) increases rapamycin (Rapa) solubility, suppresses tumor growth in breast cancer xenografts, and reduces side effects observed with free-drug controls. This new report significantly advances this carrier strategy by demonstrating the coassembly of two different ELP diblock copolymers containing drug-loading and tumor-targeting domains...
October 12, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28916764/rapid-and-reversible-epigenome-editing-by-endogenous-chromatin-regulators
#20
Simon M G Braun, Jacob G Kirkland, Emma J Chory, Dylan Husmann, Joseph P Calarco, Gerald R Crabtree
Understanding the causal link between epigenetic marks and gene regulation remains a central question in chromatin biology. To edit the epigenome we developed the FIRE-Cas9 system for rapid and reversible recruitment of endogenous chromatin regulators to specific genomic loci. We enhanced the dCas9-MS2 anchor for genome targeting with Fkbp/Frb dimerizing fusion proteins to allow chemical-induced proximity of a desired chromatin regulator. We find that mSWI/SNF (BAF) complex recruitment is sufficient to oppose Polycomb within minutes, leading to activation of bivalent gene transcription in mouse embryonic stem cells...
September 15, 2017: Nature Communications
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