Promoter selectivity of the Bacillus subtilis response regulator DegU, a positive regulator of the fla/che operon and sacB

<p>Abstract</p> <p>Background</p> <p>The response regulator DegU and its cognate histidine kinase DegS constitute a two-component system in the Gram-positive soil bacterium <it>Bacillus subtilis</it>. Unphosphorylated and phosphorylated forms of DegU are known to activate target gene transcription in <it>B. subtilis</it>. Although phosphorylated DegU (DegU-P) regulates more than one hundred and twenty genes, the targets of unphosphorylated DegU are unknown, except for <it>comK</it>.</p> <p>Results</p> <p>We found that the <it>fla/che </it>(flagella and chemotaxis) operon is positively regulated by unphosphorylated DegU. The effect was most prominent in a strain bearing the functional <it>swrAA </it>gene, a positive regulator of <it>fla/che</it>. Unphosphorylated DegU bound to two regions in the <it>fla/che </it>regulatory region containing an inverted repeat-like sequence that resembles the inverted repeat (IR) in the <it>comK </it>promoter. Mutational analysis revealed that positive regulation of <it>fla/che </it>by SwrAA requires DegU-binding. An analysis of the DegU-P-regulated gene <it>sacB </it>(levansucrase gene) by footprint and mutational analyses revealed that DegU-P bound to a direct repeat (DR) of the DegU-recognition motifs, which has been shown to be functional in vivo, while unphosphorylated DegU did not. These results strongly suggest that the arrangement of the DegU-binding motifs determines whether unphosphorylated DegU or DegU-P binds to the <it>sacB </it>promoter. The hypothesis was confirmed by observing <it>degS</it>-independent expression when the DR in the <it>sacB-lacZ </it>fusion was changed to an IR, suggesting that unphosphorylated DegU regulates the <it>sacB </it>promoter through the newly created IR. This was confirmed by binding of unphosphorylated DegU to the IR in the <it>sacB </it>promoter.</p> <p>Conclusion</p> <p>This study demonstrated that DegU positively regulates <it>flgB </it>and <it>sacB </it>through its binding to the promoter regions. We demonstrated that DegU-P prefers binding to DR but not to IR in the <it>sacB </it>promoter.</p

コウトウ ガッコウ ニ オケル シンガクシュウ シドウ ヨウリョウ ニ タイオウ シタ エイゴ シドウホウ

In March 2009, the Ministry of Education, Culture, Sports, Science and Technology （Mext） announced the New Course of Study for Senior High Schools, which will be implemented in 2013. The Mext changed the former subjects, Oral Communication I, Oral Communication II, English I, English II, Reading and Writing into Basic Communication English, Communication English I, Communication English II, Communication English III, English Expression I, English Expression II and English Conversation. The main points of the change are the following: A. Since the experience of using English is the aim, the classroom should be seen as a place where actual communication takes place. For this reason, classes should in principle be conducted in English. B. In order to develop integrated skills in communication through the learning of a foreign language, teaching materials should incorporate authentic language-use situations and suitable language functions, chosen in line with the objectives of each course. Topics should relate to subjects such as the everyday life, manners and customs, tales, geography, history, traditional culture, and natural science of people around the world, especially of people who use the language, and of Japanese people. The topics should be varied to suit the students\u27 stages of development, interests, and concerns. C. Grammar should be taught as supporting communication, and should be integrated effectively with language activities. D. In presenting necessary words, phrases, sentence structures, and grammatical structures, practical usage should be emphasized, rather than grammatical terminology and rules. As soon as the New Course of Study was announced, there were heated debates on conducting English classes in English. This paper aims to clarify the problems caused by the phrase \u27classes should in principle be conducted in English\u27 and what the practical problems are, and present various solutions to the problems, showing some examples of the techniques of English teaching

Bacillus subtilis YlxR, Which Is Involved in Glucose-Responsive Metabolic Changes, Regulates Expression of tsaD for Protein Quality Control of Pyruvate Dehydrogenase

Glucose is the most favorable carbon source for many bacteria, which have several glucose-responsive gene networks. Recently, we found that in Bacillus subtilis glucose induces the expression of the extracellular sigma factor genes sigX and sigM through the acetylation of CshA (RNA helicase), which associates with RNA polymerase (RNAP). We performed a transposon mutagenesis screen for mutants with no glucose induction (GI) of sigX-lacZ. While screening for such mutants, we recently found that the GI of sigX/M involves YlxR, a nucleoid-associated protein (NAP) that regulates nearly 400 genes, including metabolic genes. It has been shown that acetylated CshA positively regulates expression of ylxR-containing operon. Here, we report additional mutations in yqfO or tsaD required for the GI of sigX. YqfO contains a universally conserved domain with unknown function. YqfO and YlxR were found to regulate expression of the tsaEBD-containing operon. Mutational analysis using lacZ fusions revealed the adenine-rich cis-element for YlxR. TsaD is a component of the TsaEBD enzyme required for the synthesis of threonylcarbamoyl adenosine (t6A). The t6A modification of tRNA is universal across the three domains of life. Western blot analysis showed that the tsaD mutation in the presence of glucose reduced levels of soluble PdhA, PdhB, and PdhD, which are subunits of the pyruvate dehydrogenase complex (PDHc). This resulted in severely defective PDHc function and thus reduced concentrations of cellular acetyl-CoA, a reaction product of PDHc and plausible source for CshA acetylation. Thus, we discuss a suggested glucose-responsive system (GRS) involving self-reinforcing CshA acetylation. This self-reinforcing pathway may contribute to the maintenance of the acetyl-CoA pool for protein acetylation

エイゴ キョウシ ノ セイチョウ ニ ツナガル シエン ノ ゲンジョウ ト シエン ノ アリカタ ニ カンスル ケンキュウ 1

平成 21 年 3 月に告示された高等学校学習指導要領が， 平成 25 年度から学年進行で実施される． その学習指導要領第 2 章第 8 節外国語の第 3 款 「英語に関する各教科に共通する内容等」 の 4 に 「英語に関する各科目についてはその特質にかんがみ， 生徒が英語に触れる機会を充実するとともに， 授業を実際にコミュニケーションの場面とするため， 授業は英語で行うことを基本とする． （後略）」 とある． 現在文部科学省の調査によると高等学校では 90％以上の学校で 「英語Ⅰ」 の科目が主に日本語で教えられている． 「読んで， 訳して」 式の内容理解を中心とした授業である． 一方で， 平成 21 年告示の学習指導要領以前からすでに授業の大半を英語で行っている教師が少なからずいる． この前提に立ち， 学校教育現場でのうわさではなく実態を把握することによって， 英語教師の成長につながる効果的な支援の道を探ることが可能となる．この研究は， 先行研究や研修事例， 2011 年の中部英語教育学会において 「英語教師の成長につながる支援」 について発表された， 中学校・高等学校英語教師への意識調査による量的・質的研究， 英語教師の成長プロセスの認知的メカニズムなどについての研究から知見を得， 英語教師成長のための支援に関する現状と今後のあり方や改善などについて研究を進めている

ガッコウ インターンシップ ト キョウイク ジッシュウ ノ レンケツ ソノ コウカ ニ ツイテ ノ コウサツ

The aim of this paper is to introduce how the Faculty provides its students with a program of a school internship as one of the subjects for the teacher-training course and to consider the effectiveness of a link between internships at schools and teaching practice in terms of some reflections on the internships from the students and the teaching practice. A questionnaire survey was conducted in order to show how effective the internships were to the students\u27 growth and how helpful the experience of the school internships was for their teaching practice. The result describes that the subject of the school internship is completely important and indispensable for the student self-development and the assessment of their suitability for a school teacher

Differences in Ocular Complications Between Candida albicans and Non-albicans Candida Infection Analyzed by Epidemiology and a Mouse Ocular Candidiasis Model

Objectives:Candida species are a major cause of hospital infections, including ocular candidiasis, but few studies have examined the propensities of specific species to invade the eye or the unique immunological responses induced. This study examined the frequency and characteristics of species-specific Candida eye infections by epidemiology and experiments using a mouse ocular candidiasis model.Methods: We reviewed medical records of candidemia patients from January 2012 to March 2017. We also evaluated ocular fungal burden, inflammatory cytokine and chemokine profiles, and inflammatory cell profiles in mice infected with Candida albicans, Candida glabrata, or Candida parapsilosis.Results: During the study period, 20 ocular candidiasis cases were diagnosed among 99 candidemia patients examined by ophthalmologists. Although C. parapsilosis was the most frequent candidemia pathogen, only C. albicans infection was significantly associated with ocular candidiasis by multivariate analysis. In mice, ocular fungal burden and inflammatory mediators were significantly higher during C. albicans infection, and histopathological analysis revealed invading C. albicans surrounded by inflammatory cells. Ocular neutrophil and inflammatory monocyte numbers were significantly greater during C. albicans infection.Conclusion:Candida albicans is strongly associated with ocular candidiasis due to greater capacity for invasion, induction of inflammatory mediators, and recruitment of neutrophils and inflammatory monocytes