• 제목/요약/키워드: tubulin

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Heterotrimeric Kinesin 2 모터 단백질의 Carboxyl-말단과 β2-tubulin의 결합 (The Carboxyl-terminal Tail of a Heterotrimeric Kinesin 2 Motor Subunit Directly Binds to β2-tubulin)

  • 정영주;박성우;김상진;이원희;김무성;엄상화;석대현
    • 생명과학회지
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    • 제29권3호
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    • pp.369-375
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    • 2019
  • 미세소관은 알파- 와 베타-tubulin의 이량체가 종합되어 형성되며, 또한 세포내에서 tubulin 수송은 섬모나 편모와 같은 세포 부착 기관의 성장에 중요한 역할을 한다. Kinesin 2는 kinesin superfamily (KIF)의 분자 모터 단백질의 한 종류로 미세소관를 따라 다양한 운반체를 운반하며, 2종류의 다른 모터 단백질(KIF3A, KIF3B)과 kinesin-associated protein 3 (KAP3)로 구성되어 있다. Kinesin 2는 KIF3A의 cargo binding domain을 통하여 다양한 단백질과의 결합이 알려져 있지만, 아직 결합단백질의 다수는 아직 밝혀지지 않았다. 본 연구에서 KIF3A와 결합하는 단백질을 분리하기 위하여 효모 two-hybrid system을 사용하여 탐색한 결과 미세소관의 단위체의 한 종류인 ${\beta}2-tubulin$ type (Tubb2)을 분리하였다. Tubb2는 KIF3A와 결합하지만, KIF3B, KIF5B와 kinesin light chain 1 (KLC1)과는 결합하지 않았다. Tubb2의 C-말단은 KIF3A와의 결합에 필요하며, 다른 KIF3A는 Tubb의 isoforms인 Tubb1, Tubb2, Tubb3, Tubb4, Tubb5와도 결합하였다. 그러나 Tuba1은 KIF3A와 결합하지 않았다. 생쥐의 뇌 파쇄액을 KIF3A 항체로 면역침강한 결과 Tubb2는 heterotrimeric kinesin 2의 구성단백질들과 같이 침강하였다. 이러한 결과들은 heterotrimeric kinesin 2는 tubulin과 결합하여 세포 내에서 tubulins을 운반하는 것을 시사한다.

Alteration of chromosomal structure within .betha.-Tubulin and flagellar calmodulin genes during differentiation of naegleria gruberi amebae into flagellates

  • Bok, Jin-Woong;Lee, Joo-Hun
    • Journal of Microbiology
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    • 제33권3호
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    • pp.222-227
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    • 1995
  • We have examined DNase I sensitivity of .betha.-tubulin and flagellar calmodulin genes which are transiently and coordinately activated differentiation of Naegleria gruberi amebae into flagellates. The DNase I sensitivity of .betha.-tubulin and flagellar calmodulin genes changed in parallel with the changes in transcriptional activity of the respective genes during differentiation. The two genes were resistant to DNase I inamebae stage when transcription of the two genes was inactive. Forthy minutes after initiation of differentiation, when the two genes were most actively being transcribed, the two genes showed the highest sensitsivity to DNase I. One hundred and twenty minutes after initiation, the differentiation was completed and transcriptional activity of the two genes decreased to a low level. At this stage, the two genes were resistant to DNase I treatment like the ones at the amebae stage. This change in the DNase I sensitivity of the two genes was not observed when transcription of the two genes was blocked by adding cycloheximide at the beginning of differentiation.

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참나무류에 시들음병을 일으키는 Raffaelea quercus-mongolicae와 R. quercivora의 유전적 특성 (Genotypic Characterization of Oak Wilt Pathogen Raffaelea quercus-mongolicae and R. quercivora Strains)

  • 서상태;김경희;이상현;권용남;신창훈;김혜정;이상용
    • 식물병연구
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    • 제16권3호
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    • pp.219-223
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    • 2010
  • 최근 신갈나무를 중심으로한 참나무과 나무에 시들음병 피해가 증가하고 있다. 한국과 일본에서 분리한 38개의 시들음병 균주(Raffaelea quercus-mongolicae, R. quercivora)에 대해 $\beta$-tubulin 유전자 염기서열 분석과 RAPD 분석을 이용해 유전적 특성을 조사하였다. $\beta$-tubulin 유전자 염기서열을 이용한 cluster 분석결과 시들음병 균주들은 4개의 그룹으로 나뉘었으며, cluster 2와 4에는 1균주를 제외하고 모두 일본균주가 속해 있었다. RAPD 분석결과 시들음병 균주들은 3개의 그룹으로 나뉘었으며, 한국균주와 일본균주는 쉽게 구별되었다. $\beta$-tubulin 유전자 염기서열 분석과 RAPD 분석결과 한국균주와 일본균주는 상당히 다른 유전적 특성을 가지고 있었다.

Potassium Acetate Blocks Clostridium difficile Toxin A-Induced Microtubule Disassembly by Directly Inhibiting Histone Deacetylase 6, Thereby Ameliorating Inflammatory Responses in the Gut

  • Lu, Li Fang;Kim, Dae Hong;Lee, Ik Hwan;Hong, Ji;Zhang, Peng;Yoon, I Na;Hwang, Jae Sam;Kim, Ho
    • Journal of Microbiology and Biotechnology
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    • 제26권4호
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    • pp.693-699
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    • 2016
  • Clostridium difficile toxin A is known to cause deacetylation of tubulin proteins, which blocks microtubule formation and triggers barrier dysfunction in the gut. Based on our previous finding that the Clostridium difficile toxin A-dependent activation of histone deacetylase 6 (HDAC-6) is responsible for tubulin deacetylation and subsequent microtubule disassembly, we herein examined the possible effect of potassium acetate (PA; whose acetyl group prevents the binding of tubulin to HDAC-6) as a competitive/false substrate. Our results revealed that PA inhibited toxin A-induced deacetylation of tubulin and recovered toxin A-induced microtubule disassembly. In addition, PA treatment significantly decreased the production of IL-6 (a marker of inflamed tissue) in the toxin A-induced mouse enteritis model. An in vitro HDAC assay revealed that PA directly inhibited HDAC-6-mediated tubulin deacetylation, indicating that PA acted as a false substrate for HDAC-6. These results collectively indicate that PA treatment inhibits HDAC-6, thereby reducing the cytotoxicity and inflammatory responses caused by C. difficile toxin A.

Localization of cytoskeletal proteins in Pneumocystis carinii by immuno-electron microscopy

  • Yu, Jae-Ran;Pyon, Jae-Kyong;Seo, Min;Jung, Byung-Suk;Cho, Sang-Rock;Lee, Soon-Hyung;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • 제39권1호
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    • pp.13-21
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    • 2001
  • Pneumocystis carinii causes serious pulmonary infection in immuno-suppressed patients. This study was undertaken to observe the cytoskeletal proteins of P. carinii by immune-electron microscopy. P. carinii infection was experimentally induced by immunosuppression of Sprague-Dawley rats for seven weeks, and their lungs were used for the observations of this study. The gold particles localized actin, tropomyosin, and tubulin. The actin was irregularly scattered in the cytoplasm of the trophic forms but was much more concentrated in the inner space of the cell wall of the cystic forms called the inner electron-lucent layer No significant amount of tropomyosin was observed in either trophic forms or cystic forms. The tubulin was distributed along the peripheral cytoplasm and filopodia of both the trophic and cystic forms rather than in the inner side of the cytoplasm. Particularly, in the cystic forms, the amount of tubulin was increased and located mainly in the inner electron-lucent layer of the cell wall where the actin was concentrated as well. The results of this study showed that the cell wall of P carinii cystic forms is a structure whose inner side is rich in actin and tubulin. The location of the actin and tubulin in P. carinii suggests that the main role of these proteins is an involvement in the protection of cystic forms from the outside environment by maintaining rigidity of the cystic forms.

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생쥐 무릎관절 공간 발생에 있어 아포프토시스 세포에 관한 면역전자현미경적 연구 (An Immune-Electron Microscopic Study of the Apoptotic Cell during Mouse Knee Joint Development)

  • 채희선;김경용;이원복;임형수;황덕호;장가용
    • Applied Microscopy
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    • 제28권1호
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    • pp.107-119
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    • 1998
  • This study was designed to investigate the appearence and the characteristics of the apoptotic cells and the process of the joint cavity formation in mouse knee joint. Fetal mouse knee joints from 15 to 19 days of gestation were used. Paraffin-embedded serial sections, stained with H & E for light microscopic observation, Epon 812 embedded thin sections for electron microscopic observation and Lowicryl HM 20 embedded thin sections for immune-electron microscopic observation were prepared. Monoclonal antibodies to $\beta-tubulin$ and polyclonal antibodies to tissue transglutaminase were used for immune-electron microscopic study. The results obtained were as follows. 1. At 15 days of gestation, blood vessels, which have invaded in the mesenchymal cells, were present in the synovium, to form the joint cavity in the future. 2. At 16 days of gestation, the joint cleft was first appeared and several RBCs were present in the joint cleft. The invasion of blood vessels into the joint cleft was continuing, and apoptotic cells were present in the inner cell layer, adjacent to the joint cleft. Necrotic cells were also present in the outer cell layer; they were present 18 days of gestation, but apoptotic cells did not appear after 17 days of gestation. 3. In the apoptotic cells, transglutaminase were localized around vacuoles and the marginal site of the cytoplasm. 4. In the apoptotic cells, tubulin was around the endoplasmic reticulum and the marginal site of the cytoplasm. In the late stage of apoptotic cells, tubulin was localized diffusely in the cytoplasm. Tubulin was also strongly labeled around in the cytoplasm of the neighboring cell at which the apoptotic body was phagocytosed. Tubulin labeled particles were apparently increased in the seperated apoptotic bodies. On the basis of the above findings, it is proposed that during the development of the mouse knee joint, blood vessel invasion first occurs and then apoptosis and cell necrosis follow it. In the apoptotic cell, present in the synovium of the developing knee joint of the mouse. it is suggested that the redistribution of tubulin is associated with apoptotic process. And transglutaminase overexpressed in the apoptotic cell.

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Microtubule Inhibitory Effects of Various SJ Compounds on Tissue Culture Cells

  • Lee Jong Han;Kang Dong Wook;Kwon Ho Suk;Lee Sun Hwan;Park Si Kyung;Chung Sun Gan;Chon Eui Hwan;Paik Soon Young;Lee Joo Hun
    • Archives of Pharmacal Research
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    • 제27권4호
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    • pp.436-441
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    • 2004
  • SJ compounds (SJ8002 and related compounds) are a group of novel anticancer agents (Cho, Chung, Lee, Kwon, Kang, Joo, and Oh. PCT/KR02/00392). To explore the anticancer mechanism of these compounds, we examined the effect of SJ8002 on microtubules of six human cell lines. At a high concentration ($2\;{\mu}g/mL$), SJ8002 effectively disrupted microtubules of the six cell lines within 1 h. At lower concentrations ($0.05\~1.0\;{\mu}g/mL$), the antimicrotubule activity of SJ8002 varied defending on cell lines. The inhibition of in vitro polymerization of pure tubulin by SJ8002 suggested that SJ8002 acts on free tubulin, inhibits the polymerization of tubulin dimer into microtubules, and hence induces the depolymerization of microtubules.

Taxonomy of $Ganoderma$ $lucidum$ from Korea Based on rDNA and Partial ${\beta}$-Tubulin Gene Sequence Analysis

  • Park, Young-Jin;Kwon, O-Chul;Son, Eun-Suk;Yoon, Dae-Eun;Han, Woo-Ri-Ja-Rang;Yoo, Young-Bok;Lee, Chang-Soo
    • Mycobiology
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    • 제40권1호
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    • pp.71-75
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    • 2012
  • In the present study, a phylogenetic analysis was undertaken based on the internal transcribed spacer (ITS) rDNA and partial ${\beta}$-tubulin gene sequence of the $Ganoderma$ species. The size of the ITS rDNA regions from different $Ganoderma$ species varied from 625 to 673 bp, and those of the partial ${\beta}$-tubulin gene sequence were 419 bp. Based on the results, a phylogenetic tree was prepared which revealed that Korean $Ganoderma$ $lucidum$ strains belong in a single group along with a $G.$ $lucidum$ strain from Bangladesh.

송사리 모델계에서 다이아지논 노출에 대한 생물 지표로서 알파 튜블린의 동정 (The Identification of Alpha-Tubulin as a Biomarker for Diazinon Exposure in Medaka Fish)

  • 김우근;이성규;전대수;고성철;김정상
    • Environmental Analysis Health and Toxicology
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    • 제22권4호
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    • pp.305-312
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    • 2007
  • 환경오염을 신속하게 모니터링하기 위한 생물지표의 개발은 증가하고 있는 오염의 심각성에 비추어 매우 중요한 과제로 여겨지고 있다. 본 연구에서는 독성물질처리에 의하여 선택적으로 발현이 조절되는 단백질의 동정을 통하여 독성물질에 대한 단백질 생물지표를 발굴하고자 시도하였다. 즉, 송사리(Oryzias latipes)를 유기인계 살충제인 다이아지논(diazinon)에 0, 0.1, 1, 5 mg/L 농도로 24시간 노출시킨 후, 머리와 몸통부분으로 나누어 단백질 발현패턴을 분석하였다. 본 시스템에서 다이아지논 처리에 의하여 유의적으로 발현이 증가된 단백질로서 alpha-tubulin, ribonuclease pancreatic precursor, protein hfq 등을 동정하였으며, 이 가운데 alpha-tubulin과 $hsp90{\beta}$의 발현이 다이아지논 농도에 의존적으로 증가하는 것을 semi-quantitative RT-PCR방법으로 확인하였다. 이와 같이 다이아지논 처리에 특이적으로 발현이 증가된 송사리 단백질들은 노출평가를 위한 생물지표로서 개발에 응용될 수 있을 것으로 평가된다.

U-937 세포에서 이온화 방사선의 조사선량에 따른 감수성 유전자들의 발현 변화 (The mRNA Expression of Radio-Sensitive Genes Exposed to Various Dosage of Ionizing Radiation in U-937 Cell)

  • 김종수;임희영;오연경;김인규;강경선;윤병수
    • Toxicological Research
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    • 제20권1호
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    • pp.21-29
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    • 2004
  • We used cDNA microarray to assess gene expression profiles in hematopoetic cell line, U-937, exposed to low doses of ionizing irradiation. The 1,000 DNA elements on this array were PCR-amplified cDNAs selected from named human cancer related genes. According to the strength of irradiation, the levels of some gene expression were increased or decreased as dose-dependent manner. The gene expressions of Tubulin alpha, protein kinase, interferon-alpha, -beta, -omega receptor and ras homolog gene family H were significantly increased. Especially, Tubulin gene was shown 2.5 fold up-regulated manner under stress of 500 rad irradiation than 200 rad. On the other hand, fibroblast growth factor 12 and four and a half LIM domains, etc. were significantly down-regu-lated. Also, tumor protein 53(TP53) related genes that p53 inducible protein, tumor protein 53-binding protein looks of little significance as radiation sensitive manner. The radio-sensitivity of tubulin gene etc. that we proposed could be useful to rapid and correct survey for the bio-damage by exposure to low dose irradiation.