• Molecules and Cells
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• 제39권9호
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• pp.669-679
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• 2016

N-acetyl-D-glucosamine kinase (GlcNAc kinase or NAGK) primarily catalyzes phosphoryl transfer to GlcNAc during amino sugar metabolism. Recently, it was shown NAGK interacts with dynein light chain roadblock type 1 (DYNLRB1) and upregulates axo-dendritic growth, which is an enzyme activity-independent, non-canonical structural role. The authors examined the distributions of NAGK and NAGK-dynein complexes during the cell cycle in HEK293T cells. NAGK was expressed throughout different stages of cell division and immunocytochemistry (ICC) showed NAGK was localized at nuclear envelope, spindle microtubules (MTs), and kinetochores (KTs). A proximity ligation assay (PLA) for NAGK and DYNLRB1 revealed NAGK-dynein complex on nuclear envelopes in prophase cells and on chromosomes in metaphase cells. NAGK-DYNLRB1 PLA followed by Lis1/NudE1 immunostaining showed NAGK-dynein complexes were colocalized with Lis1 and NudE1 signals, and PLA for NAGK-Lis1 showed similar signal patterns, suggesting a functional link between NAGK and dynein-Lis1 complex. Subsequently, NAGK-dynein complexes were found in KTs and on nuclear membranes where KTs were marked with CENP-B ICC and nuclear membrane with lamin ICC. Furthermore, knockdown of NAGK by small hairpin (sh) RNA was found to delay cell division. These results indicate that the NAGK-dynein interaction with the involvements of Lis1 and NudE1 plays an important role in prophase nuclear envelope breakdown (NEB) and metaphase MT-KT attachment during eukaryotic cell division.

• Molecules and Cells
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• 제38권10호
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• pp.876-885
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• 2015

N-acetyl-D-glucosamine kinase (NAGK) plays an enzyme activity-independent, non-canonical role in the dendritogenesis of hippocampal neurons in culture. In this study, we investigated its role in axonal development. We found NAGK was distributed throughout neurons until developmental stage 3 (axonal outgrowth), and that its axonal expression remarkably decreased during stage 4 (dendritic outgrowth) and became negligible in stage 5 (mature). Immunocytochemistry (ICC) showed colocalization of NAGK with tubulin in hippocampal neurons and with Golgi in somata, dendrites, and nascent axons. A proximity ligation assay (PLA) for NAGK and Golgi marker protein followed by ICC for tubulin or dynein light chain roadblock type 1 (DYNLRB1) in stage 3 neurons showed NAGK-Golgi complex colocalized with DYNLRB1 at the tips of microtubule (MT) fibers in axonal growth cones and in somatodendritic areas. PLAs for NAGK-dynein combined with tubulin or Golgi ICC showed similar signal patterns, indicating a three way interaction between NAGK, dynein, and Golgi in growing axons. In addition, overexpression of the NAGK gene and of kinase mutant NAGK genes increased axonal lengths, and knockdown of NAGK by small hairpin (sh) RNA reduced axonal lengths; suggesting a structural role for NAGK in axonal growth. Finally, transfection of 'DYNLRB1 (74-96)', a small peptide derived from DYNLRB1's C-terminal, which binds with NAGK, resulted in neurons with shorter axons in culture. The authors suggest a NAGK-dynein-Golgi tripartite interaction in growing axons is instrumental during early axonal development.

• 한국자기공명학회논문지
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• 제14권2호
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• pp.127-133
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• 2010

Redox-dependent conformational change of human 8-kDa Dynein light chain (LC8) plays important role in regulating NF-${\kappa}B$ signaling pathway. In this study we characterized the structural states of the oxidized and reduced forms of LC8 by using NMR spectroscopy. The $^1H-^{15}N$ 2D HSQC spectra of oxidized LC8 indicated that no significant change in tertiary structure of LC8 occurred upon oxidation. The chemical shift perturbations of LC8 upon oxidation suggest a redox-dependent quaternary structural change.

• Journal of Chest Surgery
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• 제21권2호
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• pp.383-388
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• 1988

Immotile cilia syndrome is a congenital structural abnormality of cilia. The structural abnormality is lack of dynein arm or defective radial spoke or microtubular transposition. In this syndrome, ciliary movement is completely absent or dyskinetic and half of this syndrome shows Kartagener`s triad. We report a 13-year-old girl who had immotile cilia syndrome with Kartagener`s triad. She had been suffering from frequent respiratory infection, hemoptysis, large amount of sputum, and sinusitis. Bronchography revealed tubular bronchiectasis in right lower lobe and that lobe was resected for treatment of bronchiectasis. Histological examination of resected bronchus showed chronic bronchiectasis and electronmicroscopically complete lack of both inner and outer dynein arms. Hospital course was uneventful and symptoms were much improved.

• Tuberculosis and Respiratory Diseases
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• 제53권4호
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• pp.457-462
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• 2002

저자들은 우연히 발견된 Kartagener 증후군의 삼주징을 가진 여자환자에서 기관지점막의 섬모에 대한 전자현미경적 미세구조 분석을 시행하여 한 명의 환자에서 내, 외측 dynein arm의 결함, radial spoke의 결함, microtubule의 전위, 거대섬모 등 네 가지 이상의 다양한 미세구조결함이 혼재하는 비교적 드문 환자를 경험하였기에 보고하는 바이다.

• 한국수산과학회지
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• 제36권5호
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• pp.480-485
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• 2003

Morphology of the spermatozoa from the testes of the catfish (Pseudobagrus brevicorpus) was studied by transmission and scanning electron microscopy. The spermatozoa of P. brevicorpus are approximately $82.25\pm0.06\;{\mu}m$ in length and relatively simple cells composed of a spherical head, a short midpiece and a tail as in most teleost fish, The nucleus measuring about $2.00\pm0.02\;{\mu}m$ in length is depressed with a deep nuclear fossa of about $1.05\pm0.03\;{\mu}m$ in length three fifths of the nuclear length. The nuclear fossa contains the proximal and distal centrioles. The two centrioles are oriented approximately $150^{\circ}$ to each other. The mitochondria are arranged in two layers and their number is 12 or more. They are separated from the axoneme by the cytoplasmic canal. The axoneme is the 9+2 microtubular pattern and has inner but no outer dynein arms as in other bagrids. The axonemal fins were the closed to axonemal doublet 3 and 8. The axonemal fins and lost outer dynein arm are shared in Bagridae and the deep nuclear fossa is shared in Siluriformes. The axonemal fins observed in Bagridae and Amblycipitidae of Siluriformes might be the apomorphic character in Ostariophysi.

• Journal of Animal Science and Technology
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• 제50권6호
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• pp.849-856
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• 2008

본 연구에서는 복분자(라즈베리)의 돼지 비장 임파구에 대한 면역 반응을 조사하였다. 복분자의 70% 에틸 알코올 추출물을 돼지 비장 임파구에 처리하였다. 복분자의 추출물은 비장 면역세포의 증식을 촉진하였으며, 복분자(라즈베리)의 추출물은 돼지 비장 임파구에 대해 CD3-T 세포, CD4-T 세포와 B 세포의 구성분을 증가시켰다. 복분자 추출물은 비장 임파구 세포의 활력을 증가시킴으로써 면역반응을 향상시켰다. 본 연구에서 우리는 돼지 비장세포에 복분자 추출물을 처리한 결과 8개의 유전자 발현이 증가되었음을 확인했는데, 이들 중에는 세포구조와 면역반응에 관여하는 유전자가 포함되었다. 특히 microtubule-associated protein 4, cytoplasmic dynein heavy chain, tumor necrosis factor alpha, 및 lymphotoxin-beta receptor precursor 유전자 발현이 증가되었다. 한편 10개의 유전자는 복분자 추출물에 의해 그 발현이 감소되었다.

• Parasites, Hosts and Diseases
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• 제56권1호
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• pp.81-86
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• 2018

Four isoforms of calcium binding proteins containing 2 EF hand motifs and a dynein light chain-like domain in the human liver fluke Opisthorchis viverrini, namely OvCaBP1, 2, 3, and 4, were characterized. They had molecular weights of 22.7, 21.6, 23.7, and 22.5 kDa, respectively and showed 37.2-42.1% sequence identity to CaBP22.8 of O. viverrini. All were detected in 2- and 4-week-old immature and mature parasites. Additionally, OvCaBP4 was found in newly excysted juveniles. Polyclonal antibodies against each isoform were generated to detect the native proteins in parasite extracts by Western blot analysis. All OvCaBPs were detected in soluble and insoluble crude worm extracts and in the excretory-secretory product, at approximate sizes of 21-23 kDa. The ion-binding properties of the proteins were analyzed by mobility shift assays with the divalent cations $Ca^{2+}$, $Mg^{2+}$, $Zn^{2+}$, and $Cu^{2+}$. All OvCaBPs showed mobility shifts with $Ca^{2+}$ and $Zn^{2+}$. OvCaBP1 showed also positive results with $Mg^{2+}$ and $Cu^{2+}$. As tegumental proteins, OvCaBP1, 2, and 3 are interesting drug targets for the treatment of opisthorchiasis.

• Animal cells and systems
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• 제10권4호
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• pp.227-231
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• 2006

Micro-deletions at specific loci of the Y chromosome have been observed frequently in male infertility patients, suggesting that genes in these regions are involved in male germ cell development. DAZ is a representative male infertility gene at the AZFc locus of the Y chromosome. Since DAZ contains an RNA binding motif along with so-called a DAZ domain, it was proposed to participate in RNA metabolism during spermatogenesis. A mouse gene homologous to the human DAZ gene has been cloned and named Dazl (DAZlike). Dazl is autosomal and expressed in the testis and also at a low level in the ovary. Male mice homozygous for the Dazl null allele have small testes with a few spermatogonia and almost complete absence of germ cells beyond the spermatogonial stage, suggesting the requirement of Dazl for entry or progression through meiosis. However, its exact cellular functions have not been understood yet. In order to investigate cellular functions of Dazl, we decided to isolate candidate interacting protein genes of the mouse Dazl, using yeast two-hybrid screening. A number of candidate Dazlinteracting proteins have been isolated, such as Bprp, Acf, Hgs, Murr1, Nbak3 and Ranbp9, but dynein light chain 1 (Dlc1) was most predominant. A strong interaction of Dazl with Dlc1 suggests that Dazl might function as an mRNA adaptor to the dynein motor complex.

• 생명과학회지
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• 제33권7호
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• pp.531-537
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• 2023

세포 내 수송 및 축삭 수송은 kinesin 및 cytoplasmic dynein과 같은 미세소관 의존성 모터단백질에 의해 운반된다. Kinesin은 미세소관을 따라 미세소관의 플러스 쪽 끝으로 이동하고, dynein은 미세소관의 마이너스 쪽 끝으로 이동한다. Kinesin-1은 kinesin superfamily protein (KIF)중에서 처음으로 확인된 kinesin으로, 카복실(C)-말단 영역과 cargo간 결합을 통해 세포내 소기관, 신경전달물질 수용체 및 mRNA-단백질 복합체를 포함한 다양한 cargo의 세포내 수송 기능을 수행한다. Kinesin-1은 다양한 cargo들을 수송하지만, kinsin-1과 cargo 사이를 매개하는 어댑터/스캐폴더 단백질은 아직 완전히 확인되지 않았다. KIF5A의 C-말단 영역과 상호 작용하는 어댑터 단백질을 규명하기 위해 효모 2-하이브리드 스크리닝을 하여, cyclin-dependent kinase 2-associated protein 1 (CDK2AP1)를 확인하였다. CDK2AP1은 KIF5A의 C-말단 영역에 결합하고 KIF3A, KIF5B, KIF5C 및 kinesin light chain 1 (KLC1)과는 결합하지 않았다. CDK2AP1의 C-말단 영역은 KIF5A와의 결합에 필수적이었다. HEK-293T 세포에 CDK2AP1 및 kinesin-1은 동시 발현하여 면역침강하면 CDK2AP1 및 kinesin-1은 같이 면역침강하였다. 그리고 CDK2AP1 및 kinesin-1은 세포내에서도 같은 위치에 발현하였다. 이러한 결과들은 KIF5A-CDK2AP1결합은 kinesin-1이 cargo를 운반할 때 kinesin-1과 cargo 사이를 연결하는 어댑터 단백질 역할을 시사한다.