1,723,445 research outputs found
ASNA-1 is expressed in head neurons.
No full textRepresentative images of 1-day old adult worms expressing ASNA-1::mNG::AID (syb2249) in head neurons (white arrows). (TIFF)</p
ASNA-1 is broadly expressed in <i>C</i>. <i>elegans</i>.
No full textRepresentative fluorescence and differential interference contrast (DIC) images worms expressing ASNA-1::mNeonGeen::AID (syb2249). White arrows indicate different structures where ASNA-1::mNneonGreen::AID is present.</p
Representative fluorescence images of wild-type, <i>asna-1(ok938)</i>, <i>asna-1(ΔHis164)</i> and <i>asna-1(A63V)</i> adult worms.
No full textRepresentative fluorescence images of wild-type, asna-1(ok938), asna-1(ΔHis164) and asna-1(A63V) adult worms expressing (A) DAF-16::GFP or (B) DAF-28::GFP. White arrows indicate coelomocyte expressing DAF-28::GFP and white triangles indicate neurons expressing DAF-28::GFP. (TIFF)</p
ASNA-1 Activity Modulates Sensitivity to Cisplatin
No full textAbstract
Cancer can be cured by platinum-based chemotherapy, but resistance is a major cause of treatment failure. Here we present the nematode Caenorhabditis elegans as a model to study interactions between the platinum drug cisplatin and signaling pathways in vivo. Null mutation in a single gene, asna-1, makes worms hypersensitive to cisplatin. The metalloregulated ATPase ASNA-1 promotes insulin secretion and membrane insertion of tail-anchored proteins. Using structural data from ASNA-1 homologues, we identify specific ASNA-1 mutants that are sensitive to cisplatin while still able to promote insulin signaling. Mutational analysis reveals that hypersensitivity of ASNA-1 mutants to cisplatin remains in absence of CEP-1/p53 or apoptosis. Human ASNA1 can substitute for the worm gene, indicating a conserved function. Cisplatin sensitivity is not affected by decreased insulin signaling in wild-type nematodes or restored insulin signaling in asna-1 mutants. These findings provide a functional insight into ASNA-1, demonstrate that C. elegans can be used to characterize cisplatin resistance mechanisms, and suggest that rationally designed drugs against ASNA-1 can sensitize cancer cells to cisplatin. Cancer Res; 70(24); 10321–8. ©2010 AACR.</jats:p
Asna al-Mathaalib : syarah raudha al-thaalib
No full textKitab Asna al-Mathalib Syarh Rawdh al-Thalib merupakan sebuah kitab fiqh mazhab al-Syafi?i. Kitab Asna al-Mathalib ini disusun sebagai huraian (syarah) bagi kitab Rawdh al-Thalib karya al-?Allamah Ismail bin Abu Bakr Abdullah al-Muqri al-Yamani (837H) yang terkenal dengan gelaran Ibn al-Muqri
bi-cistronic ASNA-1^SL2::mNeonGreen::H2B.
No full text(A) Schematic representation of bi-cistronic ASNA-1^SL2::mNeonGreen::H2B (syb5730). (B) Representative fluorescence and differential interference contrast (DIC) images worms expressing ASNA-1^SL2::mNeonGreen::H2B (syb5730). (TIFF)</p
Characterization of ASNA-1 with Arsenic in Caenorhabditis elegans
No full text砷是一種自然存在的毒性物質，當暴露於砷環境的情況下會造成各種器官的病變，目前被認定為已知的人類致癌物。不論是在真核生物或原核生物，各個生物系統中都可以發現對於砷的毒性解毒機制。在Caenorhabditis elegans (C. elegans) 中,已經找到了在細菌中扮演砷解毒機制基因ArsA的同源基因asna-1。然而目前與asna-1交互作用並扮演排除三價砷的傳輸蛋白仍然未被找出。因此，本研究運用了生物資訊的方法預測出了多個可能與asna-1交互作用的基因，並且利用RNA interference (RNAi)的方法進一步篩選出此傳輸蛋白。同時，本研究也繼續探討了asna-1在C. elegans中的於砷存在時的調控機制。研究利用RNAi 篩選方法，找出了陰離子傳輸蛋白ABTS-1可能為排出砷的傳輸蛋白。我們更進一步地利用將abts-1基因移除的突變線蟲做砷的毒性測試，研究結果顯示了abts-1 缺失的線蟲對於砷的抗性有明顯的降低，證實了ABTS-1對於砷的傳輸扮演了重要的角色。研究結果亦顯示, asna-1缺失的線蟲對砷的抗性有明顯的降低。另一方面，大量表達ASNA-1蛋白質的C. elegans則可以增加對砷的抗性，更進一步證實了ASNA-1為C. elegans抗砷機制的重要蛋白。除此之外，我們也利用transgenic C. elegans以及 Real-time RT-PCR的方法對於asna-1於不同幼蟲時期的基因表達以及其暴露於砷情況下的基因調控作進一步的瞭解。我們發現了在L1幼蟲時期以及成蟲時期asna-1對於砷的誘導情況最為明顯。同時本研究也探討了asna-1以及abts-1對於C. elegans壽命的影響。結果顯示了缺乏abts-1的功能會導致C. elegans壽命的延長。本研究結果可推測出基因abts-1所轉譯的ABTS-1在對於細胞體內砷的排出扮演了重要角色並且可能為與ASNA-1作用的傳輸蛋白。同時也可推斷ASNA-1於C. elegans的幼蟲時期以及成蟲時期扮演重要的角色。Arsenic is a known human carcinogen and a potent toxin. It is a major environmental pollutant. Nearly in every organism, from prokaryotes to eukaryotes, have mechanisms for arsenic detoxification. In Caenorhabditis elegans, the gene asna-1 has been identified as the homolog of ArsA in Escherichia coli (E. coli) and involved in arsenic detoxification. To identify genes interacting with asna-1, the candidate genes were identified by gene interaction predicted program. RNA interference (RNAi) analysis was performed to screen for asna-1 interacted gene in the presence of As(III) and we found the anion transporter ABTS-1. Toxicity tests showed that both asna-1 mutant and abts-1 mutant were hypersensitive to As(III), indicating that both asna-1 and abts-1 are required for As(III) detoxification. ASNA-1 developmental mRNA expression induced by As(III) in vivo and in vitro were examined by transgenic C. elegans and real-time RT-PCR analysis, respectively. The results showed that ASNA-1 mRNA expression varied in different developmental stages of worms and adulthood had higher expression level. In addition, ASNA-1 mRNA expression was induced by As(III). Life span assay showed overexpression of ASNA-1 protein did not affect the longevity of nematodes but the deletion of abts-1 extended the life span of C. elegans. In this study, our results showed that ASNA-1 was involved in As(III) resistance and ABTS-1 might act as the corresponding transporter to As(III) detoxification. ABTS-1 might be involved in insulin-like signaling (IIS) pathway because of the extension of life span in abts-1 mutant.TABLE OF CONTENTS謝.................... .........................I文摘要...............................................IIIBSTRACT................................................VABLE OF CONTENTS................................VIIIST OF TABLES....................................IXIST OF FIGURES...........................................XIST OF APPENDIX.........................................XIBBREVIATIONS...........................................XIIHAPTER 1 INTRODUCTION..........................................1.1 Arsenic (As)....................................1.2 Detoxification system of arsenic in organisms........3.2.1 Bacteria....................................3.2.2 Saccharomyces cerevisiae.......................................4.2.3 Mammalian systems............................6.2.4 Caenorhabditis elegans (C. elegans)...........7.3 C. elegans as an animal model to explore toxicology...8.4 ArsA in organisms.............................11.4.1 ArsA in Prokaryotes..........................12.4.2 ArsA in Eukaryotes.................................12.5 Purpose of study.............................15hapter 2 MATERIALS AND METHODS.........................17.1 Chemicals................................................17.2 Strains, clone, and culture condition...............17.3 Culture and isolation of C. elegans exposed to As(III)................................................18.4 RNA Interference ( RNAi )..........................18.5 Toxicity analysis....................................19.6 Real-time RT-PCR....................................20.7 Expression of transgenic C. elegans................22.8 Arsenic exposure assay.........................22.9 Life span assay................................23.10 Statistical analysis................................25HAPTER 3 RESULTS..................................................26.1 Prediction of asna-1 interacted genes...........26.2 Identification of asna-1 interacted genes via RNA interference analysis............................27.2.1 asna-1 (RNAi)............................27.2.2 Screening of asna-1 interacted genes by RNA interference...30.3 Lethality tests of metalloids for C. elegans.........31.3.1 Effect of As(III) and Sb(III) on asna-1........31.3.2 Effect of As(III), Sb(III), and other heavy metals on abts-1...36.4 Life span assay.................................37.5 Expression of transgenic C. elegans..................41.6 Analysis of ASNA-1 mRNA expression level affected by As(III).......43hapter 4 DISCUSSION AND CONLUSION.....................49.1 Characterization of asna-1 in C. elegans.............49.2 ASNA-1 mRNA expression affected by As(III) exposure..50.3 Identification of ASNA-1 interacted genes.........52.4 Regulation of asna-1 and abts-1.................54eferences ......5
Analysis of ASNA-1 million mutation point mutants.
No full text(A) Multiple sequence alignment of ASNA-1/GET3/TRC40 comparing particular domains of the protein in 4 different species. The essential domains are highly conserved. Amino acids mentioned throughout the paper are marked in blue. (B) Confocal imaging merge of 1-day old adults co-expressing mCherry::SP12 with either ASNA-1::GFP or ASNA-1A63V::GFP. (C) Expression from the hsp-4p::GFP reporter (zcIs4) imaged by fluorescence microscopy in the 1-day old adult wild-type and asna-1(A63V) animals. hsp-4p::GFP expression quantification in the wild-type (n = 5) and asna-1(A63V) animals (n = 5). Statistical significance was determined by the independent two-sample t-test. Bars represent mean ± SD. (D) Relative mRNA analysis of ER stress reporter hsp-4 in 1-day old adult asna-1(A63V) animals. Statistical significance was determined by the independent two-sample t-test. Experiments were performed in triplicate. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. Relative mRNA analysis of (E) the mitochondrial stress reporters (hsp-6 and hsp-60) and (F) oxidative stress reporters (gst-4, gst-30, and gst-38) in 1-day old adult wild-type and asna-1(A63V) animals. Statistical significance was determined by the independent two-sample t-test. Experiments were performed in triplicate. F44B9.5 was used as a normalizing control. Bars represent mean ± SEM. (TIFF)</p
<i>asna-1(A63V)</i> mutants are defective for TAP insertion but have normal insulin secretion.
No full text(A) Representative confocal images of asna-1(+), and asna-1(A63V) 1-day-old adult animals co-expressing GFP::SEC-61β and mCherry::SP12 in the intestine. Imaging was performed in int8 and int9 cells. Scale: 10 μm and 60 μm for magnification. (B) Pearson’s correlation analysis of GFP::SEC-61β and mCherry::SP12 co-localization in asna-1(+) (n = 13) and asna-1(A63V) (n = 19). The box plot represents the average Pearson correlation coefficient (R) of the indicated strains. Statistical significance was determined by the Mann-Whitney test. (C) Band intensity quantification of membrane/cytosolic fraction of ASNA-1::GFP and ASNA-1A63V::GFP based on the western blots presented in S11 Fig. Statistical significance was determined by the independent two-sample t-test. Error bars represent ± SD. (D) Representative western blot after reducing and non-reducing SDS-PAGE to detect levels of oxidized and reduced ASNA-1A63V::GFP. Control worms expressed the ASNA-1(+)::GFP transgene. Blots were probed with anti:GFP antibody and tubulin was the loading control. For full uncropped blot source images including replicates used for quantification, see S11 Fig. Graph represents the band intensity quantification of oxidized/reduced ASNA-1 A63V::GFP. Statistical significance was determined by the independent two-sample t-test. Experiments were performed in triplicate. Bars represent ± SD. (E) Representative western blot after reducing and non-reducing SDS-PAGE to detect levels of oxidized and reduced ASNA-1A63V::GFP upon cisplatin (+CP) treatment (300 μg/mL for 6h). Control worms expressed the ASNA-1A63V::GFP transgene. Blots were probed with anti:GFP antibody and tubulin was the loading control. For full uncropped blot source images including replicates used for quantification, see S13 Fig. Graph represents the band intensity quantification of oxidized/reduced ASNA-1A63V::GFP. Statistical significance was determined by the independent two-sample t-test. Experiments were performed in triplicate. Bars represent ± SD.</p
Glycosylation of <i>asna-1(+)</i> and <i>asna-1(A63V)</i> animals.
No full textFull uncropped image of Western blot following reducing SDS-PAGE to detect glycosylated (gSEC-61β) and non-glycosylated (SEC-61β) SEC-61β in strains carrying 3xFlag::SEC-61β::opsin (rawEx64) transgene in asna-1(+) and asna-1(A63V) background. The dotted line represents a place of membrane cut in order to simultaneously probe with anti:Flag and anti:tubulin antibodies. Band intensity quantification of glycosylated vs non-glycosylated SEC-61β (gSEC-61β/ SEC-61β). Bars represent mean ± SD. Statistical significance was determined by the independent two-sample t-test. (TIFF)</p
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