• Title/Summary/Keyword: KatC

Search Result 14, Processing Time 0.028 seconds

Two Kinesins from Arabidopsis, KatB and KatC, Have a Second Microtubule-binding Site in the Tail Domain

  • Jiang, Shiling;Li, Ming;Xu, Tao;Ren, Dongtao;Liu, Guoqin
    • BMB Reports
    • /
    • v.40 no.1
    • /
    • pp.44-52
    • /
    • 2007
  • Kinesins, as a kind of microtubule-based motor proteins, have a conserved microtubule-binding site in their motor domain. Here we report that two homologous kinesins in Arabidopsis thaliana, KatB and KatC, contain a second microtubule-binding site in their tail domains. The prokaryotic-expressed N-terminal tail domain of the KatC heavy chain can bind to microtubules in an ATP-insensitive manner. To identify the precise region responsible for the binding, a serious of truncated KatC cDNAs encoding KatC N-terminal regions in different lengths, KatC1-128, KatC1-86, KatC1-73 and KatC1-63, fused to Histidine-tags, were expressed in E. coli and affinity-purified. Microtubule cosedimentation assays show that the site at amino acid residues 74-86 in KatC is important for microtubule-binding. By similarity, we obtained three different lengths of KatB N-terminal regions, KatB1-384, KatB1-77, and KatB1-63, and analyzed their microtubule-binding ability. Cosedimentation assays indicate that the KatB tail domain can also bind to microtubules at the same site as and in a similar manner to KatC. Fluorescence microscopic observations show that the microtubule-binding site at the tail domain of KatB or KatC can induce microtubules bundling only when the stalk domain is present. Through pull-down assays, we show that KatB1-385 and KatC1-394 are able to interact specifically with themselves and with each other in vitro. These findings are significant for identifying a previously uncharacterized microtubule-binding site in the two kinesin proteins, KatB and KatC, and the functional relations between them.

Mutations of katG and inhA in MDR M. tuberculosis (국내에서 분리된 다제 내성 결핵균의 katG 와 inhA 변이 다양성 및 그 빈도)

  • Lin, Hai Hua;Kim, Hee-Youn;Yun, Yeo-Jun;Park, Chan Geun;Kim, Bum-Joon;Park, Young-Gil;Kook, Yoon-Hoh
    • Tuberculosis and Respiratory Diseases
    • /
    • v.63 no.2
    • /
    • pp.128-138
    • /
    • 2007
  • Backgrounds: Mutations of katG and inhA (ORF and promoter) are known to be related to isoniazid (INH) resistance of Mycobacterium tuberculosis. Because reports on these mutations in Korean isolates are limited (i.e. only the frequency of katG codon 463 was evaluated.), we tried to know the kinds of mutations of two genes and their frequencies in INH resistant Korean M. tuberculosis strains. Methods: PCR was performed to amplify katG (2,223 bp), inhA ORF (-77~897, 975 bp), and inhA promoter (-168~80, 248 bp) from 29 multidrug resistant M. tuberculosis (MDR-TB) DNAs prepared by bead beater-phenol method. Their sequences were determined and analyzed by ABI PRISM 3730 XL Analyzer and MegAlign package program, respectively. Results: All of the isolates had more than one mutation in katG or inhA gene. Twenty seven (93%) of 29 tested strains had katG mutations, which suggests that katG is a critical gene determining INH resistance of M. tuberculosis. Amino acid substitutions, such as Arg463Leu and Ser315Thr, due to point mutations of the katG were the most frequent (62.1% and 55.2%) mutations. In addition, deletion of the katG gene was frequently observed (17.2%). Analyzed Korean MDR-TB isolates also had variable inhA mutations. Point mutation of inhA promoter region, such as -15 ($C{\rightarrow}T$) was frequently found. Substitution of amino acid (Lsy8Asn) due to point mutation ($AAA{\rightarrow}AAC$) of inhA ORF was found in 1 isolate. Interestingly, 14 point mutated types that were not previously reported were newly found. While four types resulted in amino acid change, the others were silent mutations. Conclusions: Although it is not clear that the relationship of these newly found mutations with INH resistance, they show marked diversity in Korean MDR-TB strains. It also suggests their feasibility as a molecular target to supplement determining the INH resistance of clinical isolates because of the possible existence of low-level INH resistant strains.

A neonate with Say-Barber-Biesecker-Young-Simpson syndrome with a novel pathogenic mutation in KAT6B gene: A case report

  • Shin, Ji Hye;Lim, Han Hyuk;Gang, Mi Hyeon;Kim, Seon Young;Yang, Shin-seung;Chang, Mea-young
    • Journal of Genetic Medicine
    • /
    • v.18 no.2
    • /
    • pp.147-151
    • /
    • 2021
  • The Say-Barber-Biesecker-Young-Simpson variant of Ohdo syndrome (SBBYSS) (Online Mendelian Inheritance in Man #603736) is a rare autosomal dominant disorder and clinically features blepharophimosis with ptosis, a mask-like facial appearance, cryptorchidism, congenital heart defect, long thumbs/great toes, and thyroid dysfunction. The etiology of SBBYSS has been shown to be due to heterozygous KAT6B gene mutation. Here we report a case of a neonate with SBBYSS identified a novel mutation in KAT6B gene. The patient showed typical dysmorphic facies, cryptorchidism with micropenis, overriding fingers, and long thumbs and toes at birth. He had also hypothyroidism, large atrial septal defect, and sensorineural hearing loss. The next generation sequencing identified a heterozygous novel variant, c.5206C>T (p.Gln1736Ter) in KAT6B gene. At the 9 months of age, he underwent patch closure for atrial septal defect. Until the 12-month follow-up, he was under-developed.

Rapid Drug Susceptibility Testing for Isoniazid and Rifampicin by Reverse Hybridization Assay (역교잡반응법을 이용한 아이소니아지드 및 리팜피신 신속감수성검사)

  • Park, Young Kil;Yu, Hee Kyoung;Ryu, Sung Weon;Bai, Gill Han
    • Tuberculosis and Respiratory Diseases
    • /
    • v.55 no.5
    • /
    • pp.440-448
    • /
    • 2003
  • Background : Development of rapid drug susceptibility testing provides the opportunity for rapid identification of individuals with drug resistant tubercle bacilli, allowing selection of appropriate therapeutic regimens. Methods : A total of 502 drug resistant isolates were subjected to reverse blot hybridization assay to detect mutations within genes (rpoB, katG, inhA, and ahpC) associated with rifampicin (RMP) and isoniazid (INH) resistance. Results : Among the 264 RMP resistant strains ($RMP^R$) tested, the most prevalent mutation was the Ser531Leu seen in 121 strains (46%). The second common mutation occurred in 84 strains (32%) at codon 526. And 27 strains (10%) showed the mutation at codon 516. Among all 469 INH resistant strains ($INH^R$), the katG mutation was responsible for INH. The inhA mutation was present in 88 strains (19%). In 11 isolates (2%), coexisting of the katG and inhA mutations were identified. Reverse hybridization assay successfully detected over 80% of $INH^R$ and over 92% of $RMP^R$ among Korean isolates. CONCLUSION: Reverse hybridization was useful for rapid detection of $INH^R$ and $RMP^R$.

Molecular Analysis of Isoniazid-Resistance Related Genes of Mycobacterium tuberculosis Isolated from Korea

  • Hwang Joo Hwan;Jeong Eun Young;Choi Yeon Im;Bae Kiho;Song Taek Sun;Cho Sang-Nae;Lee Hyeyoung
    • Biomedical Science Letters
    • /
    • v.11 no.4
    • /
    • pp.455-463
    • /
    • 2005
  • Resistance to isoniazid (INH), which is one of the most important drugs in Mycobacterium tuberculosis chemotherapy, has been associated with mutations in genes encoding the mycobacterial catalse-peroxidase (katG), the enoyl acyl carrier protein (ACP) reductase (inhA), alkyl hydroperoxide reductase (ahpC), beta-ketoacyl acyl carrier protein synthase (kasA), and NADH dehydrogenase (ndh). In this study, we examined INH-resistance related genes in 50 INH-resistant and 24 INH-susceptible isolates by PCR-sequence analysis. In brief, mutations at the katG gene were found at codon 315 alone (2/50), at codon 463 alone (19/50), and both at 315 and 463 (29/50). However, while mutations at codon 315 were only detected in INH-resistant isolates, mutations at codon 463 were also detected in INH-susceptible isolates indicating mutations at 463 alone do not seem to confer resistance to INH. Similar to the case of katG 463, some of inhA mutations were also found among INH-susceptible isolates. For example, whereas mutations at 8 upstream of the start codon (UPS) and 15 UPS of the inhA gene were detected only in INH-resistant isolates, mutations at 101, 115, and 125 UPS were detected only in INH-susceptible isolates. Many different kinds of mutations were detected in INH­resistant isolates at ahpC, oxyR gene, and intergenic region of the oxyR-ahpC genes. Howerver, the mutations were not found oxyR and the intergenic regions in INH-susceptible isolates. No mutations were found at either kasA or at ndh gene among INH-resistant isolates. In conclusion, some of mutations such as katG 315, inhA promotor region, and oxyR-ahpC seem to be strongly related to INH-resistance. Currently we are developing a molecular diagnostic method based on these results.

  • PDF

An Ecological Study on the Process of Plant Community Formation in Tidal Land (간척지 식물군락형성 과정에 관한 연구 -목포지방을 중심으로-)

  • 김철수
    • Journal of Plant Biology
    • /
    • v.14 no.4
    • /
    • pp.27-33
    • /
    • 1971
  • Information this research an attempt was made to investigate the plant community structure and the process of its formation in the tidal area surrounding Mokpo City the examined area included the stand in Sam-Hak Do wherein sands have inflowed, and the stand in Kat-Ba-Woo which has been left as tidal soil land. Two hundred stands were sampled by making 100m line transects 5m apart and establishing 30cm$\times$30cm quadrates 10m apart on each tape. These data were used to establish frequency, cover degree, density, standing crop (g/$m^2$), contained Cl in the soil, and pH. Results of the research are as following: 1. There were 25 species of plants in the stands from Sam-Hak Do, 18 in the stands from Kat-Ba-Woo, and so 28 species in all. Regardings the importance value, the rank of dominant species is Salicornia herbacea L., Suaeda maritima Dumorties, Diplachne fusca L., Echinochola hispidula Nakai, Cyperus iria L., Setaria lutescens Hubbard in Sam-Hak Do, and Suaeda maritima Dumorties, Echinochloa hispidula Nakai, Aster tripolium L., Scripus maritimus L., Salicornia herbacea L., Diplachne fusca L., in the lands from Kat-Ba-Woo. Among them are five kinds of halophytes Salicornia herbacea L., Suaeda maritima Dumorties, Atriplex Gmelini C.A. Meyer, Aster tripolium L., and Limonium tetragonum Bullock; and two Kinds of naturalized plants Diplachan fusca L., Aster subulata. M. In the stands from Sam-Hak Do there was evidence of a secondary succession phenomena in the presence of Cyperaceae such as Cyperus liria L., Juncus decipiens Nakai, and Fimbristylis longispica Steudal, which could not be found in the stands from Kat-Ba-Woo. 2. On the number of species of vegetation, the further inland from the floodgate the higher the number; that is, the lower the content of Cl the higher the number of species. 3. On the distribution of the vegetation; comparing DFD index and Cl content, the main plants are Salicornia herbacea L., Suaeda maritima Dumorties, Atriplex gmelini C.A. Meyer, Diplachne fuscal, Echinochola hispidula Nakai where the Cl content of soil is more than 13.2%. 4. On the standing crop; Salicornia herbacea L., which has high resistance to salt, was half the total was production of 1090/$m^2$ while Suaeda maritima Dumorties was 1/4, Diplachne fusca L. 1/8, and Echinochloa the least. Analyzing the result, the research indicates that the main factor in plant community formation in tidal land is tolerance for Cl, and that a plan for utilizing the halophytes which are abundant in tidal land should be devised.

  • PDF

A Study on the Protein Productivity of the Promoters for Cold Inducible Genes in Escherichia coli (대장균 저온 유도성 유전자 Promoter의 단백질 생산성에 관한 연구)

  • Kim, So-Yeon;Kim, Su-Hyun;Heo, Mi-Ae;Lee, Sun-Gu
    • KSBB Journal
    • /
    • v.21 no.6 s.101
    • /
    • pp.461-465
    • /
    • 2006
  • We investigated the protein productivity of the promoters for genes showing prolonged induction upon cold shock in Escherichia coli. Six low temperature inducible genes (frdA, glpB, hypB, katG, nupG, ompT) were selected based on the previously reported cDNA microarray based global transcription profiling of Escherichia coli Kl2 in response to cold shock. Their promoter regions were isolated from the genomic DNA of E. coli JM109 and expression levels induced by the promoters were examined by using green fluorescence protein (GFP) as a reporter at $15^{\circ}C$ and $37^{\circ}C$. Among the six promoters, the promoter for nupG showed the highest and prolonged expression at both temperatures and the cold inducibility of nupG promoter was not observed.

Treatment of Human Thyroid Carcinoma Cells with the G47delta Oncolytic Herpes Simplex Virus

  • Wang, Jia-Ni;Xu, Li-Hua;Zeng, Wei-Gen;Hu, Pan;Rabkin, Samuel D.;Liu, Ren-Rin
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.16 no.3
    • /
    • pp.1241-1245
    • /
    • 2015
  • Background: Thyroid carcinoma is the most common malignancy of the endocrine organs. Although the majority of thyroid cancer patients experience positive outcomes, anaplastic thyroid carcinoma is considered one of the most aggressive malignancies. Current therapeutic regimens do not confer a significant survival benefit, and new therapies are urgently needed. Oncolytic herpes simplex virus (oHSV) may represent a promising therapy for cancer. In the present study, we investigated the therapeutic effects of a third-generation HSV vector, $G47{\Delta}$, on various human thyroid carcinoma cell lines in vitro. Two subcutaneous (s.c.) models of anaplastic thyroid carcinoma were also established to evaluate the in vivo anti-tumor efficacy of $G47{\Delta}$. Materials and Methods: The human thyroid carcinoma cell line ARO, FRO, WRO, and KAT-5, were infected with $G47{\Delta}$ at different multiplicities of infection (MOIs) in vitro. The survival rates of infected cells were calculated each day. Two s.c. tumor models were established using ARO and FRO cells in Balb/c nude mice, which were intratumorally (i.t.) treated with either $G47{\Delta}$ or mock. Tumor volumes and mouse survival times were documented. Results: $G47{\Delta}$ was highly cytotoxic to different types of thyroid carcinomas. For ARO, FRO, and KAT-5, greater than 30% and 80% of cells were killed at MOI=0.01 and MOI=0.1, respectively on day 5. WRO cells displayed modest sensitivity to $G47{\Delta}$, with only 21% and 38% of cells killed. In the s.c. tumor model, both of the anaplastic thyroid carcinoma cell lines (ARO and FRO) were highly sensitive to $G47{\Delta}$; $G47{\Delta}$ significantly inhibited tumor growth and prolonged the survival of mice bearing s.c. ARO and FRO tumors. Conclusions: The oHSV $G47{\Delta}$ can effectively kill different types of human thyroid carcinomas in vitro. $G47{\Delta}$ significantly inhibited growth of anaplastic thyroid carcinoma in vivo and prolonged animal survival. Therefore, $G47{\Delta}$ may hold great promise for thyroid cancer patients.

The Implementation of SEED Cipher Algorithm Test Module Applied CMVP Test (CMVP 테스트를 적용한 SEED 암호 알고리즘 모듈 구현)

  • Park, Seong-Gun;Jeong, Seong-Min;Seo, Chang-Ho;Kim, Il-Jun;Shin, Seung-Jung;Kim, Seok-Woo
    • Proceedings of the Korea Information Processing Society Conference
    • /
    • 2003.05c
    • /
    • pp.1937-1940
    • /
    • 2003
  • 정보보호 평가는 크게 시스템 평가인 CC(Common Criteria)평가와 암호모듈 평가인 CMVP(Cryptographic Module Validation Program)평가로 나눌 수 있다. 본 논문은 국내 표준 암호 알고리즘 SEED를 북미의 CMVP의 3가지 블록 알고리즘 시험방법인 KAT(Known Answer Test), MCT(Monte C미개 Test), MMT(Multi-block Message Test)를 JAVA환경에 적용하여 시범 구현하였다. 테스트 방법으로 CMVP의 MOVS, TMOVS, AESAVS를 선정하여 FIPS 표준을 적용하였다. 구현 환경으로는 JCE기반의 Cryptix를 채택하여 CMVP의 블록 암호 알고리즘 테스트 시스템 중 일부를 구현하였다.

  • PDF

Cellular Responses to Alcohol in Escherichia coli, Clostridium acetobutylicum, and Saccharomyces cerevisiae (알코올에 대한 Escherichia coli, Clostridium acetobutylicum, Saccharomyces cerevisiae의 반응)

  • Park, Ju-Yong;Hong, Chun-Sang;Han, Ji-Hye;Kang, Hyun-Woo;Chung, Bong-Woo;Choi, Gi-Wook;Min, Ji-Ho
    • Korean Chemical Engineering Research
    • /
    • v.49 no.1
    • /
    • pp.105-108
    • /
    • 2011
  • The increased concern for the security of the oil supply and the negative impact of fossil fuels on the environment, particularly greenhouse gas emissions, has put pressure on society to find renewable fuel alternatives. Compared to the traditional biofuel, ethanol, higher alcohols offer advantage as gasoline substitutes because of their higher energy density and lower hygroscopicity. For this reason, microbial fermentation is known as potential producers for sustainable energy carriers. In this study, bacterial responses including cellular and molecular toxicity were studied in three different microorganisms, such as Escherichia coli, Clostridium acetobutylicum, and Saccharomyces cerevisiae. In this study, it was analyzed specific stress responses caused by ethanol and buthanol using four different stress responsive genes, i.e. fabA, grpE, katG and recA. The expression levels of these genes were quantified by semi-quantitative reverse transcription-PCR. It was found that four genes have shown different responsive patterns when E. coli cultures were under stressful conditions caused by ethanol and buthanol, respectively. Therefore, in this study, the stress responsive effects caused by these alcohols and the extent of each stress response can be analyzed using the expression levels and patterns of different stress responsive genes.