• Title/Summary/Keyword: CP1 domain

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Structural characterization of the putative DNA-binding domain of CP2c and its relevance to zinc binding

  • Ryu, Ki-Sung;Jo, Ku-Sung;Kim, Na-Young;Jeon, Eun-Jae;Park, Sung Jean;Kim, Hyun-Hwi;Kim, Eun-Hee;Kim, Chan-Gil;Kim, Chul Geun;Won, Hyung-Sik
    • Journal of the Korean Magnetic Resonance Society
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    • v.23 no.1
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    • pp.20-25
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    • 2019
  • The transcription factor CP2c has been recently validated as an oncogenic protein that can serve as a promising target for anticancer therapy. We have recently documented that a recombinant protein corresponding to the putative DNA-binding region (residues 63-244) of CP2c adopted two different conformers, one of which is dominated by zinc binding. However, in the present study, a longer construct encompassing residues 63-302 appeared to form a single structural domain. This domain could be considered to adopt a functionally relevant fold, as the known specific binding of a dodecapeptide to this protein was evident. Hence, the residues 63-302 region rather than 63-244 can be regarded as a natively folded structural domain of CP2c. In addition, it was confirmed that zinc ions can bind to this putative DNA-binding domain of CP2c, which resulted in reduced stability of the protein. In this context, it is suggested that the mode of action of CP2c would resemble that of tumor suppressor p53.

Structural assessment of the tetramerization domain and DNA-binding domain of CP2c

  • Jo, Ku-Sung;Ryu, Ki-Sung;Yu, Hee-Wan;Lee, Seu-Na;Kim, Ji-Hun;Kim, Eun-Hee;Wang, Chae-Yeon;Kim, Chan-Gil;Kim, Chul Geun;Won, Hyung-Sik
    • Journal of the Korean Magnetic Resonance Society
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    • v.22 no.4
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    • pp.119-124
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    • 2018
  • Although the transcription factor CP2c has been recently validated as a promising target for development of novel anticancer therapy, its structure has not been solved yet. In the present study, the purified recombinant protein corresponding to the tetramerization domain of CP2c appeared to be well folded, whereas the Elf-1 domain showed a largely unfolded conformation. Particularly, the Elf-1 domain, which contains the putative DNA-binding region, showed a conformational equilibrium between relatively less-ordered and well-ordered conformers. Interestingly, addition of zinc shifted the equilibrium to the relatively more structured conformer, whereas zinc binding decreased the overall stability of the protein, leading to a promoted precipitation. Likewise, a dodecapeptide that has been suggested to bind to the Elf-1 domain also appeared to shift the conformational equilibrium and to destabilize the protein. These results constitute the first structural characterization of the CP2c domains and newly suggest that zinc ion might be involved in the conformational regulation of the protein.

The coat protein of Turnip crinkle virus is required a full-length to maintain suppressing activity to RNA silencing but no relation with eliciting resistance by N-terminal region in Arabidopsis.

  • Park, Chang-Won;Feng Qu;Tao Ren;T. Jack Morris
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.76.1-76
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    • 2003
  • The coat protein (CP) of Turnip crinkle virus (TCV) is organized into 3 distinct domains, R domain (RNA-binding) connected by an arm, 5 domain and P domain. We have previously shown that the CP of TCV strongly suppresses RNA silencing, and have mapped N-terminal R domain of which is also the elicitor of resistance response in the Arabidopsis ecotype Di-17 carrying the HRT resistance gene. In order to map the region in the TCV CP that is responsible for silencing suppression, a series of CP mutants were constructed, transformed into Agrobacterium, coinfiltrated either with HC-Pro (the helper component proteinase of tobacco etch potyvirus) known as a suppressor of PTGS or GFP constructs into leaves of Nicotiana benthmiana expressing GFP transgenically. In the presence of HC-Pro, all CP mutants were well protected, accumulating mutant CP mRNAs and their proteins even 5 days post-infiltration (DPI). In the presence of GFP, some mutant constructs which showed the accumulation of CP mutants and GFP mRNAs at early stage but eventually degraded at 5 DPI. Only a mutant which carrying 4 amino acid deletion of R domain was tolerable to maintain suppressing activity, suggesting that the suppressing activity is not directly related with the eliciting activity. A transient assay also revealed that the mutants synthesized their proteins, suggesting that a full length of CP sequences and its intact structure are required to stabilize CP, which suppresses the RNA silencing.

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DGNSS-CP Performance Comparison of Each Observation Matrix Calculation Method (관측 행렬 산출 기법 별 DGNSS-CP 성능 비교)

  • Shin, Dong-hyun;Lim, Cheol-soon;Seok, Hyo-jeong;Yoon, Dong-hwan;Park, Byungwoon
    • Journal of Advanced Navigation Technology
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    • v.20 no.5
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    • pp.433-439
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    • 2016
  • Several low-cost global navigation satellite system (GNSS) receivers do not support general range-domain correction, and DGNSS-CP (differential GNSS) method had been suggested to solve this problem. It improves its position accuracy by projecting range-domain corrections to the position-domain and then differentiating the stand-alone position by the projected correction. To project the range-domain correction, line-of-sight vectors from the receiver to each satellite should be calculated. The line-of-sight vectors can be obtained from GNSS broadcast ephemeris data or satellite direction information, and this paper shows positioning performance for the two methods. Stand-alone positioning result provided from Septentrio PolaRx4 Pro receiver was used to show the difference. The satellite direction information can reduce the computing load for the DGNSS-CP by 1/15, even though its root mean square(RMS) of position error is bigger than that of ephemeris data by 0.1m.

Three Common Subunits in the Editing Domains of Class Ia tRNA Synthetases.

  • Lee, Keun-Woo;Kwon, Yong-Jung;Briggs, James M.
    • Journal of Industrial Technology
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    • v.24 no.B
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    • pp.139-142
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    • 2004
  • To identify conserved structural or functional subunit(s) in the CP1 (editing) domains of class Ia tRNA synthetases, five available structures were compared and analyzed. Through sequence alignments of the CP1 domains, three conserved regions were found near the amino acid binding site in the editing domain. Structural overlapping of the three subunits clearly showed that there exist three common structural subunits in all of the five editing RS structures. The new alignment suggests a translocation movement of the CP1 domain caused by the binding with tRNA. Based on the experimental and modeling results, it is proposed that subunits 1 and 3 accommodate the incoming amino acid binding, while subunit 2 contributes to the interactions with the adenosine ring of the A76 to stabilize the overall tRNA binding.. Since these subunits are critical for the editing reaction, we expect that these key structures should be conserved through all class Ia editing RSs.

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Intrinsically disordered fold of a PIAS1-binding domain of CP2b

  • Jo, Ku-Sung;Jo, Hae-Ri;Kim, Chul Geun;Kim, Chan-Gil;Won, Hyung-Sik
    • Journal of the Korean Magnetic Resonance Society
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    • v.18 no.1
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    • pp.30-35
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    • 2014
  • The transcription factor CP2 regulates various biological systems at diverse tissues and cells. However, none of the four CP2 isoforms has been solved in structure yet. In particular, two different regions of the CP2b isoform have been characterized to interact with the PIAS1 in nucleus to regulate the ${\alpha}$-globin gene expression. Among them, in this study, the region encompassing residues 251-309 of CP2b was prepared as a recombinant protein and its solution structure was characterized by NMR spectroscopy. The results indicated that the CP2b(251-309) fold belongs to typical IDRs (intrinsically disordered regions), likely to facilitate promiscuous interactions with various target proteins. Unfortunately, however, its interaction with the N-terminal domain of PIAS1 (residues 1-70), which has been identified as one of the CP2b-binding sites, was not observed in the NMR-based titration experiments. Therefore, it could be postulated that the 251-309 region of CP2b would not contact with the PIAS1(1-70), but alternatively interact with another CP2b-binding region that encompasses residues 400-651 of PIAS1.

A Feasibility Test on the DGPS by Correction Projection Using MSAS Correction

  • Yoon, Dong Hwan;Park, Byungwoon;Yun, Ho;Kee, Changdon
    • Journal of Positioning, Navigation, and Timing
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    • v.3 no.1
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    • pp.25-30
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    • 2014
  • Differential Global Positioning System-Correction Projection (DGPS-CP) algorithm, which has been suggested as a method of correcting pre-calculated position error by projecting range-domain correction to positional domain, is a method to improve the accuracy performance of a low price GPS receiver to 1 to 3 m, which is equivalent to that of DGPS, just by using a software program without changing the hardware. However, when DGPS-CP algorithm is actually realized, the error is not completely eliminated in a case where a reference station does not provide correction of some satellites among the visible satellites used in user positioning. In this study, the problem of decreased performance due to the difference in visible satellites between a user and a reference station was solved by applying the Multifunctional Transport Satellites (MTSAT) based Augmentation System (MASA) correction to DGPS-CP, instead of local DGPS correction, by using the Satellite Based Augmentation System (SBAS) operated in Japan. The experimental results showed that the accuracy was improved by 25 cm in the horizontal root mean square (RMS) and by 20 cm in the vertical RMS in comparison to that of the conventional DGPS-CP.

Three Common Subunits in Editing Domains of Class Ia tRNA Synthetases

  • Lee, Keun-Woo;Kwon, Yong-Jung;Briggs, James M.
    • Bulletin of the Korean Chemical Society
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    • v.28 no.2
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    • pp.207-210
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    • 2007
  • To identify structural or functional common subunit(s) in the CP1 (editing) domains of class Ia tRNA synthetases, five available structures were compared and analyzed. Through the sequence alignments and structural overlapping of the CP1 domains, three conserved regions were identified near the amino acid binding site in the editing domain. Structural overlapping of the three subunits clearly showed the existence of three common structural subunits in all of the five editing RS structures. Based on the established experimental results and our modeling results, it is proposed that subunits 1 and 3 accommodate the incoming amino acid binding, while subunit 2 contributes to the interactions with the adenosine ring of the A76 to stabilize the overall tRNA binding. Since these subunits are critical for the editing reaction, we expect that these key structures should be conserved through the most class Ia editing RSs.

Nucleotide Polymorphisms of Cytochrome P450 Genes in Domestic Garlic Cultivars (국내 재배마늘의 Cytochrome P450 유전자의 염기다형성 분포)

  • Kwon, Soon-Tae;Chung, Jinbo
    • Korean Journal of Plant Resources
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    • v.31 no.5
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    • pp.531-537
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    • 2018
  • This study was carried to survey distribution of the nucleotide polymorphisms in heme-binding (HB) domain, which is highly conserved region between 1,210 and 1,240 bp of cytochrome P450, in domestic garlic cultivars. 120 garlic cultivars collected from Korea were classified into seven HB domain variation based on the nucleotide sequence of the domain. Northern type garlic cultivars, collected from Kyungpook, Chungnam, Chungpook and Kangwon province, showed 51.3% of KP2 type nucleotide sequence, 5'-TTT/GGC/GGT/GGA/CGG/AGA/ATA/TGT/CCT/GGA-3' with coding amino acid FGGGRRICPG, 13.7% of KP1, 11.3% of CP, 8.8% of CM and 5% of KW2 types. Southern type cultivars, collected from Kyungnam province, showed 52.5% of KM type nucleotide sequence, 5'-TTT/GGC/GCA/GGA/CGG/AGA/ATT/TGT/CCT/GGA-3' with coding amino acid FGAGRRICPG, 22.5% of KP2, 5.0% of KW2 and 2,5% of CP type nucleotide sequence. These results showed that Korean garlics were cultivated in highly mixed condition even in the same region.