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https://www.readbyqxmd.com/read/28815417/cellular-interactome-analysis-of-vaccinia-virus-k7-protein-identifies-three-transport-machineries-as-binding-partners-for-k7
#1
Ye Li, Youyang Ke, Leiliang Zhang
Identification of viral-host interacting proteins will contribute to understanding of how poxvirus exploits the host cellular machinery. The vaccinia virus gene K7R encodes a conserved protein K7 in most orthopoxviruses. To gain insight into the biology of K7, we investigated the cellular interactome of K7 by GST pulldown coupled with mass spectrometry. The top categories of identified proteins contained components of trafficking machineries. We selected key components of three transport machineries including coatomer, retromer, and CHEVI to further confirm their binding abilities to K7...
August 16, 2017: Virus Genes
https://www.readbyqxmd.com/read/28814803/the-bhlh-transcription-factor-cgbhlh001-is-a-potential-interaction-partner-of-cdpk-in-halophyte-chenopodium-glaucum
#2
Juan Wang, Gang Cheng, Cui Wang, Zhuanzhuan He, Xinxin Lan, Shiyue Zhang, Haiyan Lan
Plants have evolved different abilities to adapt to the ever-fluctuating environments for sessility. Calcium-dependent protein kinase (CDPK) is believed to play a pivotal role in abiotic stress signaling. So far, study on the specific substrates that CDPK recognized in response to adversity is limited. In the present study, we revealed a potential interaction between CDPK and a bHLH transcription factor under salt stress in Chenopodium glaucum. First, we identified a CgCDPK, which was up-regulated under salt and drought stress; then by Y2H screening, CgCDPK was detected to be involved in interaction with a bHLH TF (named as CgbHLH001), which also positively respond to salt and drought stress...
August 16, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28813705/the-lncrna-tug1-ezh2-axis-promotes-pancreatic-cancer-cell-proliferation-migration-and-emt-phenotype-formation-through-sponging-mir-382
#3
Liang Zhao, Hongwei Sun, Hongru Kong, Zongjing Chen, Bicheng Chen, Mengtao Zhou
BACKGROUND/AIMS: Pancreatic carcinoma (PC) is the one of the most common and malignant cancers worldwide. LncRNA taurine upregulated gene 1 (TUG1) was initially identified as a transcript upregulated by taurine, and the abnormal expression of TUG1 has been reported in many cancers. However, the biological role and molecular mechanism of TUG1 in PC still needs further investigation. METHODS: Quantitative real-time PCR (qRT-PCR) was performed to measure the expression of TUG1 in PC cell lines and tissues...
August 15, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28808999/studying-protein-protein-interactions-by-biotin-ap-tagged-pulldown-and-ltq-orbitrap-mass-spectrometry
#4
Zhongqiu Xie, Yuemeng Jia, Hui Li
The study of protein-protein interactions represents a key aspect of biological research. Identifying unknown protein binding partners using mass spectrometry (MS)-based proteomics has evolved into an indispensable strategy in drug discovery. The classic approach of immunoprecipitation with specific antibodies against the proteins of interest has limitations, such as the need for immunoprecipitation-qualified antibody. The biotin AP-tag pull-down system has the advantage of high specificity, ease of use, and no requirement for antibody...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28768865/ubiquitin-ligase-wwp1-interacts-with-ebola-virus-vp40-to-regulate-egress
#5
Ziying Han, Cari A Sagum, Fumio Takizawa, Gordon Ruthel, Corbett T Berry, Jing Kong, J Oriol Sunyer, Bruce D Freedman, Mark T Bedford, Sachdev S Sidhu, Marius Sudol, Ronald N Harty
Ebola virus (EBOV) is a member of the Filoviridae family and the cause of hemorrhagic fever outbreaks. The EBOV VP40 (eVP40) matrix protein is the main driving force for virion assembly and budding. Indeed, expression of eVP40 alone in mammalian cells results in the formation and budding of virus-like particles (VLPs), which mimics the budding process and morphology of authentic, infectious EBOV. To complete the budding process, eVP40 utilizes its PPxY L-domain motif to recruit a specific subset of host proteins containing one or more modular WW-domains that then function to facilitate efficient production and release of eVP40 VLPs...
August 2, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28759334/b-cell-tetramer-development-for-veterinary-vaccinology
#6
Michael C Rahe, Kevin L Gustafson, Michael P Murtaugh
Immunological memory is elicited after either vaccination or natural exposure to a pathogen and is essential for protection against re-exposure. Despite its critical importance, the ability to interrogate the veterinary animal memory immune response has long been hindered by a paucity of tools to assess immunological memory. As a result, the evaluation and analysis of protective immune responses that predict immune protection in food and fiber animals and facilitate vaccine development are obstructed. To fill this gap in knowledge in swine, we created a B cell tetramer to porcine reproductive and respiratory syndrome virus (PRRSV) nonstructural protein 7 (nsp7) to efficiently and effectively investigate the memory B cell response, a hallmark of anti-viral immunity...
July 31, 2017: Viral Immunology
https://www.readbyqxmd.com/read/28749462/keratin-23-promotes-telomerase-reverse-transcriptase-expression-and-human-colorectal-cancer-growth
#7
Ningning Zhang, Rui Zhang, Kun Zou, Wendan Yu, Wei Guo, Yingying Gao, Jia Li, Mei Li, Yidi Tai, Wenlin Huang, Chun Song, Wuguo Deng, Xiaonan Cui
The overexpression of human telomerase reverse transcriptase (hTERT) has been associated with the proliferation and migration of colorectal cancer (CRC) cells. We investigated the roles of KRT23 and hTERT in promoting CRC cell proliferation and migration. We verified the relationship between KRT23 and hTERT in CRC using streptavidin-agarose pulldown and chromatin immunoprecipitation (ChIP) assays. One hundred and fifty-four human CRC specimens were analyzed using immunohistochemistry. The roles of KRT23 and hTERT in cell growth and migration were studied using siRNA and lentiviruses in vivo and in vitro...
July 27, 2017: Cell Death & Disease
https://www.readbyqxmd.com/read/28733876/human-papillomavirus-type-8-e7-protein-binds-nuclear-myosin-1c-and-downregulates-the-expression-of-pre-rrna
#8
Evelyn Oswald, Eileen Reinz, Renate Voit, François Aubin, Angel Alonso, Eeva Auvinen
Our aim was to search for new cellular binding partners for the E6 and E7 oncogenes of beta human papillomaviruses (HPV), whose direct role in skin carcinogenesis has not been thoroughly investigated. By employing glutathione S-transferase pulldown and coimmunoprecipitation, we identified nuclear myosin 1c as a binding partner of HPV 8 E7 protein. As nuclear myosin 1c is an essential component of the RNA polymerase I transcription complex, we studied the effects of HPV 8 E7 protein expression on ribosomal RNA (rRNA) expression...
July 21, 2017: Virus Genes
https://www.readbyqxmd.com/read/28719045/mical2-promotes-breast-cancer-cell-migration-by-maintaining-egfr-stability-and-egfr-p38-signaling-activation
#9
Yueyuan Wang, Wenjie Deng, Yujie Zhang, Shixiu Sun, Shuo Zhao, Yan Chen, Xuyang Zhao, Lei Liu, Jun Du
AIM: MICAL2, a cytoskeleton dynamics regulator, is identified associated with survival and metastasis of several types of cancers recently. The present study was designed to investigate the role of MICAL2 in breast cancer cell migration as well as its underlying mechanisms. METHODS: The relationship between MICAL2 and EGF/EGFR signaling was analyzed by using gene overexpression and knockdown techniques. Cell migration was measured by wound healing assays. Activation of EGF/EGFR singaling pathways were evaluated by immunofluorescence, qPCR, Western blotting and zymography techniques...
July 18, 2017: Acta Physiologica
https://www.readbyqxmd.com/read/28704058/elucidating-protein-dna-interactions-in-human-alphoid-chromatin-via-hybridization-capture-and-mass-spectrometry
#10
Katherine E Buxton, Julia Kennedy-Darling, Michael R Shortreed, Nur Zafirah Zaidan, Michael Olivier, Mark Scalf, Rupa Sridharan, Lloyd M Smith
The centromere is the chromosomal locus where the kinetochore forms, and is critical for ensuring proper segregation of sister chromatids during cell division. A substantial amount of effort has been devoted to understanding the characteristic features and roles of the centromere, yet some fundamental aspects of the centromere, such as the complete list of elements that define it, remain obscure. It is well known that human centromeres include a highly repetitive class of DNA known as alpha satellite, or alphoid, DNA...
July 13, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/28701209/alantolactone-a-natural-sesquiterpene-lactone-has-potent-antitumor-activity-against-glioblastoma-by-targeting-ikk%C3%AE-kinase-activity-and-interrupting-nf-%C3%AE%C2%BAb-cox-2-mediated-signaling-cascades
#11
Xun Wang, Zhenlong Yu, Chao Wang, Wei Cheng, Xiangge Tian, Xiaokui Huo, Yan Wang, Chengpeng Sun, Lei Feng, Jinshan Xing, Yulong Lan, Dongdong Sun, Qingjuan Hou, Baojing Zhang, Xiaochi Ma, Bo Zhang
BACKGROUND: Glioblastoma multiforme (GBM) is one of the most refractory and palindromic central nervous system (CNS) neoplasms, and current treatments have poor effects in GBM patients. Hence, the identification of novel therapeutic targets and the development of effective treatment strategies are essential. Alantolactone (ATL) has a wide range of pharmacological activities, and its anti-tumor effect is receiving increasing attention. However, the molecular mechanism underlying the anti-GBM activity of ATL remains poorly understood...
July 12, 2017: Journal of Experimental & Clinical Cancer Research: CR
https://www.readbyqxmd.com/read/28699205/native-kinesin-1-does-not-bind-preferentially-to-gtp-tubulin-rich-microtubules-in-vitro
#12
Qiaochu Li, Stephen J King, Jing Xu
Molecular motors such as kinesin-1 work in small teams to actively shuttle cargos in cells, for example in polarized transport in axons. Here, we examined the potential regulatory role of the nucleotide state of tubulin on the run length of cargos carried by multiple kinesin motors, using an optical trapping-based in vitro assay. Based on a previous report that kinesin binds preferentially to GTP-tubulin-rich microtubules, we anticipated that multiple-kinesin cargos would run substantially greater distances along GMPCPP microtubules than along GDP microtubules...
July 12, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/28683446/lncrna-snhg6-is-associated-with-poor-prognosis-of-gastric-cancer-and-promotes-cell-proliferation-and-emt-through-epigenetically-silencing-p27-and-sponging-mir-101-3p
#13
Kai Yan, Jie Tian, Wenzheng Shi, Hao Xia, Yuanfang Zhu
Background/Amis: Long non-coding RNAs (lncRNAs), a novel class of transcripts, have been shown to play critical roles in diverse cellular biological processes, including tumorigenesis. Small nucleolar RNA host gene 6 (SNHG6) regulates various biological processes in cancer cells. However, the biological role of SNHG6 in gastric cancer still remains to be explored. The aim of this study is to investigate the characteristic of the SNHG6 in gastric cancer. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the expression of SNHG6 in gastric cancer tissues and cell lines...
June 27, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28657147/lncrna-pvt1-promotes-pancreatic-cancer-cells-proliferation-and-migration-through-acting-as-a-molecular-sponge-to-regulate-mir-448
#14
Liang Zhao, Hongru Kong, Hongwei Sun, Zongjing Chen, Bicheng Chen, Mengtao Zhou
The identification and characterization of long non-coding RNAs (lncRNAs) in diverse biological process has currently developed rapidly. LncRNA-PVT1, located adjacent to the MYC locus on chromosomal region 8q24, has been reported to be associated with many biological processes. However, the function and mechanism of PVT1 in pancreatic carcinoma (PC) is poorly understood. In this present study, we first measured the level of PVT1 in the PC cell lines and tissues by quantitative real-time PCR (qRT-PCR), and then employed loss-of-function and gain-of-function approaches to explore the association between PVT1 expression levels and PC cell proliferation/migration ability...
June 28, 2017: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/28636190/uch-l1-promotes-invasion-of-breast-cancer-cells-through-activating-akt-signaling-pathway
#15
Yanhong Luo, Jianfeng He, Chunlin Yang, Matthew Orange, Xingcong Ren, Nick Blair, Tao Tan, Jin-Ming Yang, Hua Zhu
As a de-ubiquitin enzyme, ubiquitin C-terminal hydrolase (UCH)-L1 has been shown to be overexpressed in several human cancers. However, the function of UCH-L1 in invasion of breast cancers is still unclear. Here we report that the expression of UCH-L1 is significantly higher in cancer cells with higher invasive ability. While ectopic UCH-L1 expression failed to alter cell proliferation in MCF-7 cells, it caused a significant upregulation of cellular invasion. Furthermore, siRNA mediated knockdown of UCH-L1 led to suppression of invasion in UCH-L1 overexpressing MCF-7 cells...
June 21, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28628370/multiple-joint-exercises-using-elastic-resistance-bands-vs-conventional-resistance-training-equipment-a-cross-over-study
#16
Vegard M Iversen, Paul Jarle Mork, Ottar Vasseljen, Ronny Bergquist, Marius S Fimland
Previous studies indicate that elastic resistance bands (ERB) can be a viable option to conventional resistance-training equipment (CRE) during single-joint resistance exercises, but their efficacy has not been established for several commonly used multiple-joint resistance exercises. Thus, we compared muscular activation levels in four popular multiple-joint exercises performed with ERB (TheraBand(®)) vs. CRE (Olympic barbell or cable pulley machines). In a cross-over design, men and women (n = 29) performed squats, stiff-legged deadlifts, unilateral rows and lateral pulldown using both modalities...
September 2017: European Journal of Sport Science
https://www.readbyqxmd.com/read/28625921/binding-of-coronin-1b-to-t%C3%AE-ri-negatively-regulates-the-tgf%C3%AE-1-signaling-pathway
#17
Geun-Young Kim, Hyun-Joung Lim, Hyun-Young Park
Coronin 1B is an actin-binding protein that regulates several actin-dependent cellular processes including migration and endocytosis. However, the role of coronin 1B in the tumor growth factor (TGF)β signaling pathway is largely unknown. Here, we investigated whether coronin 1B affects the TGFβ signaling cascade and found that coronin 1B negatively regulates the TGFβ signaling pathway. Immunoprecipitation and glutathione-S-transferase-pulldown assays revealed that coronin 1B directly associated with TGFβ receptor I (TβRI)...
August 19, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28607052/efficient-ultra-high-affinity-chromatography-in-a-one-step-purification-of-complex-proteins
#18
Marina N Vassylyeva, Sergiy Klyuyev, Alexey D Vassylyev, Hunter Wesson, Zhuo Zhang, Matthew B Renfrow, Hengbin Wang, N Patrick Higgins, Louise T Chow, Dmitry G Vassylyev
Protein purification is an essential primary step in numerous biological studies. It is particularly significant for the rapidly emerging high-throughput fields, such as proteomics, interactomics, and drug discovery. Moreover, purifications for structural and industrial applications should meet the requirement of high yield, high purity, and high activity (HHH). It is, therefore, highly desirable to have an efficient purification system with a potential to meet the HHH benchmark in a single step. Here, we report a chromatographic technology based on the ultra-high-affinity (Kd ∼ 10(-14)-10(-17) M) complex between the Colicin E7 DNase (CE7) and its inhibitor, Immunity protein 7 (Im7)...
June 27, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28592633/the-phospho-caveolin-1-scaffolding-domain-dampens-force-fluctuations-in-focal-adhesions-and-promotes-cancer-cell-migration
#19
Fanrui Meng, Sandeep Saxena, Youtao Liu, Bharat Joshi, Timothy H Wong, Jay Shankar, Leonard J Foster, Pascal Bernatchez, Ivan R Nabi
Caveolin-1 (Cav1), a major Src kinase substrate phosphorylated on tyrosine-14 (Y14), contains the highly conserved membrane-proximal caveolin scaffolding domain (CSD; amino acids 82-101). Here we show, using CSD mutants (F92A/V94A) and membrane-permeable CSD-competing peptides, that Src kinase-dependent pY14Cav1 regulation of focal adhesion protein stabilization, focal adhesion tension, and cancer cell migration is CSD dependent. Quantitative proteomic analysis of Cav1-GST (amino acids 1-101) pull downs showed sixfold-increased binding of vinculin and, to a lesser extent, α-actinin, talin, and filamin, to phosphomimetic Cav1Y14D relative to nonphosphorylatable Cav1Y14F...
August 1, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28584154/pa3225-is-a-transcriptional-repressor-of-antibiotic-resistance-mechanisms-in-pseudomonas-aeruginosa
#20
Clayton W Hall, Li Zhang, Thien-Fah Mah
The tssABC1 locus is part of the Hcp secretion island I (HSI-I) type VI secretion system (T6SS) in Pseudomonas aeruginosa Previous work implicated the tssC1 gene in P. aeruginosa biofilm-specific antibiotic resistance, and tssC1 is preferentially expressed in biofilms compared to planktonic cells. Using a DNA-dependent protein pulldown approach, we discovered that PA3225, an uncharacterized LysR-type transcriptional regulator, specifically bound to the tssABC1 upstream regulatory region. The deletion of PA3225 led to a 2-fold decrease in tssA1 expression levels in planktonic cells compared to the wild type, and tssA1 expression was slightly reduced in ΔPA3225 biofilms compared to wild-type biofilms...
August 2017: Antimicrobial Agents and Chemotherapy
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