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Molecular Insights into Toluene Sensing in the TodS/TodT Signal Transduction System
Koh, Serry,Hwang, Jungwon,Guchhait, Koushik,Lee, Eun-Gyeong,Kim, Sang-Yoon,Kim, Sujin,Lee, Sangmin,Chung, Jeong Min,Jung, Hyun Suk,Lee, Sang Jun,Ryu, Choong-Min,Lee, Seung-Goo,Oh, Tae-Kwang,Kwon, Ohsu American Society for Biochemistry and Molecular Bi 2016 The Journal of biological chemistry Vol.291 No.16
<P>TodS is a sensor kinase that responds to various monoaromatic compounds, which either cause an agonistic or antagonistic effect on phosphorylation of its cognate response regulator TodT, and controls <I>tod</I> operon expression in <I>Pseudomonas putida</I> strains. We describe a molecular sensing mechanism of TodS that is activated in response to toluene. The crystal structures of the TodS Per-Arnt-Sim (PAS) 1 sensor domain (residues 43–164) and its complex with toluene (agonist) or 1,2,4-trimethylbenzene (antagonist) show a typical β2α3β3 PAS fold structure (residues 45–149), forming a hydrophobic ligand-binding site. A signal transfer region (residues 150–163) located immediately after the canonical PAS fold may be intrinsically flexible and disordered in both apo-PAS1 and antagonist-bound forms and dramatically adapt an α-helix upon toluene binding. This structural change in the signal transfer region is proposed to result in signal transmission to activate the TodS/TodT two-component signal transduction system. Site-directed mutagenesis and β-galactosidase assays using a <I>P. putida</I> reporter strain system verified the essential residues involved in ligand sensing and signal transfer and suggest that the Phe<SUP>46</SUP> residue acts as a ligand-specific switch.</P>
Show and Tell : cell biology of pathogen invasion
Serry Koh,Shauna Somerville 한국작물학회 2007 한국작물학회 학술발표대회 논문집 Vol.2007 No.11
The powdery mildews are obligate biotrophic fungi and are one of the most economically important groups of plant pathogens (Agrios, 1997). As a family, the powdery mildews infect a broad range of plant species including barley, wheat, pea, apple, sugar beet, and grape (Braun, 1987). Powdery mildew pathogens uptake nutrients by forming a feeding structure, the haustorium, within 12‐18 hours after infection (hai) in their respective host plants. Because of the initial stages of these plant pathogen invasion are mostly confined to a limited number of host cells, it’s often too late to find out till the infections widely spread out. To identify the earliest and often transient responses to pathogen attack, there is considerable interest in monitoring the subcellular events that occur specifically in living host cells. Recent improvements in live cell imaging using fluorescent‐tagged markers have expanded the scope of experiments that can be performed. Changes in the subcellular distribution of organelles as well as fluorescently tagged proteins can be monitored in real time in living tissues during pathogen attack, and the dynamic nature of such changes across space and over time can be determined. The application of these sensitive imaging methods has extended earlier observations made with Nomarski microscopy or inferred from static transmission electron micrographs about the focal accumulation of subcellular organelles at sites of pathogen attack. In addition, recent experiments have demonstrated the focused accumulation and interaction of specific plant proteins at penetration sites, opening a new window on early host responses and raising questions about the underlying plant processes that sense and direct this marshalling of host resources to block pathogen entry.
A novel light‐dependent selection marker system in plants
Koh, Serry,Kim, Hongsup,Kim, Jinwoo,Goo, Eunhye,Kim, Yun‐,Jung,Choi, Okhee,Jwa, Nam‐,Soo,Ma, Jun,Nagamatsu, Tomohisa,Moon, Jae Sun,Hwang, Ingyu Blackwell Publishing Ltd 2011 Plant biotechnology journal Vol.9 No.3
<P><B>Summary</B></P><P>Photosensitizers are common in nature and play diverse roles as defense compounds and pathogenicity determinants and as important molecules in many biological processes. Toxoflavin, a photosensitizer produced by <I>Burkholderia glumae</I>, has been implicated as an essential virulence factor causing bacterial rice grain rot. Toxoflavin produces superoxide and H<SUB>2</SUB>O<SUB>2</SUB> during redox cycles under oxygen and light, and these reactive oxygen species cause phytotoxic effects. To utilize toxoflavin as a selection agent in plant transformation, we identified a gene, <I>tflA</I>, which encodes a toxoflavin‐degrading enzyme in the <I>Paenibacillus polymyxa</I> JH2 strain. TflA was estimated as 24.56 kDa in size based on the amino acid sequence and is similar to a ring‐cleavage extradiol dioxygenase in the <I>Exiguobacterium</I> sp. 255‐15; however, unlike other extradiol dioxygenases, Mn<SUP>2+</SUP>and dithiothreitol were required for toxoflavin degradation by TflA. Here, our results suggested toxoflavin is a photosensitizer and its degradation by TflA serves as a light‐dependent selection marker system in diverse plant species. We examined the efficiencies of two different plant selection systems, toxoflavin/<I>tflA</I> and hygromycin/hygromycin phosphotransferase (<I>hpt</I>) in both rice and <I>Arabidopsis</I>. The toxoflavin/<I>tflA</I> selection was more remarkable than hygromycin/<I>hpt</I> selection in the high‐density screening of transgenic <I>Arabidopsis</I> seeds. Based on these results, we propose the toxoflavin/<I>tflA</I> selection system, which is based on the degradation of the photosensitizer, provides a new robust nonantibiotic selection marker system for diverse plants.</P>
고세리(Serry Koh),최유리(Youri Choi),이주영(Joo Young Lee),장지영(Jiyoung Jang),최규환(Kyuwhan Choi) 한국육종학회 2021 한국육종학회지 Vol.53 No.4
New Breeding Technology (NBT) refers to gene editing technologies that are used to develop crop plants with beneficial traits,from biotic/abiotic resistance to nutritional enhancement, including zinc finger nucleases (ZFN), transcription activator-like effector nucleases,clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9, meganucleases, and oligo directed mutagenesis. A total of 1,119 valid NBT patents were analyzed in this study to examine global trends in the patent and market expansion strategies formajor patent applicants. Based on the claims specified, valid patents in each patent office were analyzed through the applicant’s country oforigin, field of technology, and plant/crop species. Patents claiming applications of CRISPR-based technology to major crop plants, includingrice, corn, wheat, tomato, and canola, have rapidly increased in the China National Intellectual Property Administration (CNIPA) since 2013. The patent family size (PFS) can be used as an indicator of intellectual property (IP)-based market expansion strategies and target marketsof interests of patent applicants. Many university- and research-oriented institution Chinese applicants showed low PFS (2.1) because theyfiled patents mostly in CNIPA. In contrast, high PFS of US and German (DE) applicants such as Corteva Agriscience (US), KWS SAATAG (DE), Cellectics (FR), and Syngenta Participations AG (CH) represented their active strategies for global gene-edited crop market expansion. Corteva Agriscience (US, 238 patents) was the global leader in patents using NBT, ranging from ZFN to CRISPR-based technologies appliedto most major crops, including corn, soybean, and wheat.
JEONG, YEONHWA,CHEONG, HOON,CHOI, OKHEE,KIM, JUNG KYU,KANG, YONGSUNG,KIM, JINWOO,LEE, SEUNGDON,KOH, SERRY,MOON, JAE SUN,HWANG, INGYU Blackwell Publishing Ltd 2011 Molecular plant pathology Vol.12 No.4
<P><B>SUMMARY</B></P><P>The host specificity of <I>Ralstonia solanacearum</I>, the causal organism of bacterial wilt on many solanaceous crops, is poorly understood. To identify a gene conferring host specificity of the bacterium, SL341 (virulent to hot pepper but avirulent to potato) and SL2029 (virulent to potato but avirulent to hot pepper) were chosen as representative strains. We identified a gene, <I>rsa1</I>, from SL2029 that confers avirulence to SL341 in hot pepper. The <I>rsa1</I> gene encoding an 11.8‐kDa protein possessed the perfect consensus hrp<SUB>II</SUB> box motif upstream of the gene. Although the expression of <I>rsa1</I> was activated by HrpB, a transcriptional activator for <I>hrp</I> gene expression, Rsa1 protein was secreted in an Hrp type III secretion‐independent manner. Rsa1 exhibited weak homology with an aspartic protease, cathepsin D, and possessed protease activity. Two specific aspartic protease inhibitors, pepstatin A and diazoacetyl‐<SMALL>d</SMALL>,<SMALL>l</SMALL>‐norleucine methyl ester, inhibited the protease activity of Rsa1. Substitution of two aspartic acid residues with alanine at positions 54 and 59 abolished protease activity. The SL2029 <I>rsa1</I> mutant was much less virulent than the wild‐type strain, but did not induce disease symptoms in hot pepper. These data indicate that Rsa1 is an extracellular aspartic protease and plays an important role for the virulence of SL2029 in potato.</P>