1,721,153 research outputs found
Functional characterization of Su(fu) phosphorylation in Drosophila.
No full text<p>(A) Hh-dependent transcriptional activation was measured by activity of a <i>ptc</i>-luciferase reporter normalized to constitutively expressed control <i>Renilla</i> luciferase. Cells were stimulated with or without conditioned medium containing HhN, the N-terminal signaling domain of the Hh protein. (B) Identification of phosphorylation sites in Su(fu). A schematic diagram of Su(fu) is shown and phosphoserine residues identified by mass spectrometry in endogenous Su(fu) purified from HhN-stimulated <i>Drosophila</i> cl-8 cells are marked with “P”. (C) Western blot analysis of Ala or Glu substitution of Su(fu) phosphoresidues. (D) Endogenous <i>Su(fu)</i> mRNA was targeted using dsRNA corresponding to the 3’ untranslated region (3’UTR), and expression constructs encoding Su(fu) but lacking the 3’UTR sequence were used to test for function of Su(fu) variants, with pathway activity measured as in (A). A representative experiment from at least three independent experiments is shown. Error bars show mean +/- standard deviation. Statistical significance was measured by Student’s <i>t</i>-test: **** (P<0.0001), *** (0.00010.05).</p
Regulation of Ci and Su(fu) nuclear import in Drosophila
No full textAbstractThe Hedgehog (Hh) signal transduction pathway plays a central role in the development of invertebrates and vertebrates. While much is known about the pathway, the role of Suppressor of fused (Su(fu)), a component of the pathway's signaling complex has remained enigmatic. Previous studies have linked Su(fu) to the cytoplasmic sequestration of the zinc finger transcription factor, Cubitus interruptus (Ci), while other studies suggest a role in modulating target gene expression. In examining the cell biology of the pathway, we have found that like its vertebrate homologue, Drosophila Su(fu) enters the nucleus. Furthermore, we find that the nuclear import of Su(fu) occurs in concert with that of Ci in response to Hh signaling. Here, we examine the mechanism by which Su(fu) regulates Ci import by investigating the importance of the Ci nuclear localization signal (NLS) and the effect of adding an additional NLS. Finally, we demonstrate that Ci can bring Su(fu) with it to a multimerized Ci DNA binding site. These results provide a basis for understanding the dual roles played by Su(fu) in the regulation of Ci
Physical and functional characterization of Su(fu) and Cos2 interactions with Ci.
No full text<p>(A) Schematic representation of Ci variants and their interactions with Su(fu) and Cos2. (B) Su(fu) represses Ci activity through binding to a.a. 1–346 of Ci. (Top) S2R+ cells were transfected with indicated expression constructs, and with dsRNA against Cos2 to deplete endogenous Cos2, which could indirectly bring V5-Su(fu) to FLAG-Ci constructs through endogenous Fu. (Bottom) Su(fu) binding is required for Ci suppression by Su(fu). Increasing amounts of V5-Su(fu) (0, 25, 50, 125 ng) and fixed amount of FLAG-Ci constructs (25 ng) were transfected in S2R+ cells with Cos2 RNAi to deplete endogenous Cos2. (C) CDN and CORD mediate most but not all of the Cos2 binding and repression of Ci. (Top) S2R+ cells were transfected with dsRNA against Su(fu) as well as indicated expression constructs, and subjected to either anti-FLAG or anti-Myc IP. (Bottom) Increasing amounts of Myc-Cos2 (0, 25, 50, 125 ng) and fixed amount of FLAG-Ci constructs (25 ng) were expressed in S2R+ cells with Su(fu) RNAi to deplete endogenous Su(fu). A representative experiment from at least three independent experiments is shown. Error bars show mean +/- standard deviation. Statistical significance was measured by Student’s <i>t</i>-test: **** (P<0.0001), *** (0.00010.05). P values in (B) and (C) were calculated for pairs of the same DNA mass of V5-Sufu (B) or Myc-Cos2 (C) construct between WT Ci-expressing cells and a Ci deletion/truncation variant-expressing cells.</p
Rab23 negatively regulates Gli1 transcriptional factor in a Su(Fu)-dependent manner
No full textHedgehog (Hh) signaling, via the key signal transducer Smoothened (SMO) and Gli transcription factors, is essential for embryonic development and carcinogenesis. While the biological relevance of hedgehog signaling to cancer is well established, very little is known about the molecular mechanisms by which signaling transduction of this pathway occurs. Rab23 was discovered as a negative regulator of the Hh pathway through a mouse genetic study. Here we report that Rab23 directly associates with Su(Fu) and inhibits Gli1 function in a Su(Fu)-dependent manner. By confocal microscope and immunoprecipitation, we detected interaction between Rab23 and Su(Fu). Using Gli1-mediated reporter gene analysis, we found that Rab23 can suppress Gli1 transcriptional activity in wild type but not Su(Fu) null fibroblasts. Similarly, Rab23 expression reduced the nuclear localization of Gli1 in wild type but not Su(Fu) null fibroblast cells. Consistent with the GTPase motif in the protein, we showed that Rab23 has GTPase activity. The dominant negative form of Rab23 was unable to suppress Gli1-mediated transcriptional activity. Taken together, these data provide evidence to support that Rab23 negatively regulates Gli1 activity in a Su(Fu)-dependent manner
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
MO Injection Experiments against Su(fu), Hip, and Ptc2
No full text<div><p>(A) Dorsal view of the eye showing the lens in the eye chamber.</p><p>(B–D) Dorsal view of embryos injected with the indicated MOs, resulting in a phenocopy of <i>dre, uki,</i> and <i>lep</i> mutants.</p><p>(E) A wild-type ear showing the presence of the dorsolateral septum (arrow), which is not present after injections with the indicated MOs (F–H, arrow).</p><p>(I) Injections with control MOs against the initiation codon of Su(fu) results in chevron-shaped somites with an angle of 97°.</p><p>(J) Injection of MOs against Su(fu) results in a more obtuse angle of the somite (126°).</p></div
Variations on the Author
Get PDF“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Get PDFWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
A Study of Rural Politics in Taiping's Su-fu Province
No full text李秀成地方建设的新思维,为苏福省地方建设规划了基本原则与发展方向。在李秀成的影响下,苏福省出现了一个以熊万荃和钱桂仁为代表的具有儒家化特征的施政群体。苏福省当局普遍实行的是"着佃交粮"与"招业收租"并举的政策。田凭政策是较"着佃交粮"更高层次的社会经济政策,虽然有很强的政治性,却丝毫不具备任何革命性变革的意义。只有1862年钟志成推行的田凭政策,才真正具有土地革命的涵义。苏福省的乡官制度,在继承前期基本功能的同时,其主观能动性更趋活跃。Li Xiu-cheng's new thought on local construction programmed basic principle and developmental circumstances for the local construction in Taiping's Su-Fu Province.Under Ii Xiu-cheng's influence,there appeared a political group characteristic of Confusionization in Su-Fu Province whose representative figures were Xiong Wan-quan and Qian Gui-ren.The Su-Fu's administration generally practised "Zhuo-Dian-Jiao-Liang" and "Zhao-Ye-Shou-Zu" policies together.Tian-Ping policy was a superior policy than "Zhuo-Dian-Jiao-Liang" policy.It had not any revolutionary significance,although it possessed strong political sense.The subjective initiative of local-official system was brought into full play in Su-fu Province
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