• Journal of Microbiology and Biotechnology
• /
• 제9권5호
• /
• pp.582-588
• /
• 1999

A Brevibacterium lactofermentum gene coding for a glucose-specific permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned, by complementing an Escherichia coli mutation affecting a ptsG gene with the B. lactofermentum genomic library, and completely sequenced. The gene was identified as a ptsG, which enables an E. coli transformant to transport non-metabolizable glucose analogue 2-deoxyglucose (2DG). The ptsG gene of B. lactofermentum consists of an open reading frame of 2,025 nucleotides encoding a polypeptide of 674 amino acid residues and a TAA stop codon. The 3' flanking region contains two stem-loop structures which may be involved in transcriptional termination. The deduced amino acid sequence of the B. lactofermentum enzyme $II^{GIe}$ specific to glucose ($EII^{GIe}$) has a high homology with the Corynebacterium glutamicum enzyme $II^{Man}$ specific to glucose and mannose ($EII^{Man}$), and the Brevibacterium ammoniagenes enzyme $II^{GIc}$ specific to glucose ($EII^{GIc}$). The 171-amino-acid C-terminal sequence of the $EII^{Glc}$ is also similar to the Escherichia coli enzyme $IIA^{GIc}$ specific to glucose ($IIA^{GIc}$). It is interesting that the arrangement of the structural domains, IIBCA, of the B. lactofermentum $EII^{GIc}$ protein is identical to that of EIIs specific to sucrose or $\beta$-glucoside. Several in vivo complementation studies indicated that the B. lactofermentum $EII^{Glc}$ protein could replace both $EII^{ Glc}$ and $EIIA^{Glc}$ in an E. coli ptsG mutant or crr mutant, respectively.

• Molecules and Cells
• /
• 제26권4호
• /
• pp.404-408
• /
• 2008

The genome of replication-competent BERV ${\gamma}4$ provirus, which is the most abundant ERV family in the bovine genome, was characterized in detail. The BERV ${\gamma}4$ genome showed that BERV ${\gamma}4$ harbors 8576 nucleotides and has the typical 5'-long terminal repeat (LTR)-gag-pro-pol-env-LTR-3' retroviral organization with a long leader region positioned before the gag open reading frame. Multiple sequences analysis showed that the nucleotide difference between 5' and 3' LTRs was 4.2% (mean value 0.042) in average, suggesting that the provirus formed at most 13.3 million years ago. Gag separated by a stop codon from pro-pol in the same reading frame, while env resides in another reading frame lacking of a functional surface domain. According to the current bovine genome sequence assembly, the full-length BERV ${\gamma}4$ provirus sequences were only found in the chromosomes 1, 2, 6, 10, 15, 23, 26, 28, X, and unassigned, although the partial sequences almost evenly distributed in the entire bovine genome. This is the first detailed study describing the genome structure of BERV ${\gamma}4$, the most abundant ERV family present in bovine genome. Combined with our recent reports on characterization of ERVs in bovine, this study will contribute to illuminate ERVs in the cattle of which no information was previously available.

• BMB Reports
• /
• 제52권7호
• /
• pp.445-450
• /
• 2019

TTYH2 is a calcium-activated, inwardly rectifying anion channel that has been shown to be related to renal cancer and colon cancer. Based on the topological prediction, TTYH2 protein has five transmembrane domains with the extracellular N-terminus and the cytoplasmic C-terminus. In the present study, we identified a vesicle transport protein, ${\beta}$-COP, as a novel specific binding partner of TTYH2 by yeast two-hybrid screening using a human brain cDNA library with the C-terminal region of TTYH2 (TTYH2-C) as a bait. Using in vitro and in vivo binding assays, we confirmed the protein-protein interactions between TTYH2 and ${\beta}$-COP. We also found that the surface expression and activity of TTYH2 were decreased by co-expression with ${\beta}$-COP in the heterologous expression system. In addition, ${\beta}$-COP associated with TTYH2 in a native condition at a human colon cancer cell line, LoVo cells. The over-expression of ${\beta}$-COP in the LoVo cells led to a dramatic decrease in the surface expression and activity of endogenous TTYH2. Collectively, these data suggested that ${\beta}$-COP plays a critical role in the trafficking of the TTYH2 channel to the plasma membrane.

• The Korean Journal of Physiology and Pharmacology
• /
• 제23권3호
• /
• pp.219-227
• /
• 2019

Polycystic kidney disease 2-like-1 (PKD2L1), polycystin-L or transient receptor potential polycystin 3 (TRPP3) is a TRP superfamily member. It is a calcium-permeable non-selective cation channel that regulates intracellular calcium concentration and thereby calcium signaling. Although the calmodulin (CaM) inhibitor, calmidazolium, is an activator of the PKD2L1 channel, the activating mechanism remains unclear. The purpose of this study is to clarify whether CaM takes part in the regulation of the PKD2L1 channel, and if so, how. With patch clamp techniques, we observed the current amplitudes of PKD2L1 significantly reduced when co-expressed with CaM and $CaM{\triangle}N$. This result suggests that the N-lobe of CaM carries a more crucial role in regulating PKD2L1 and guides us into our next question on the different functions of two lobes of CaM. We also identified the predicted CaM binding site, and generated deletion and truncation mutants. The mutants showed significant reduction in currents losing PKD2L1 current-voltage curve, suggesting that the C-terminal region from 590 to 600 is crucial for maintaining the functionality of the PKD2L1 channel. With PKD2L1608Stop mutant showing increased current amplitudes, we further examined the functional importance of EF-hand domain. Along with co-expression of CaM, ${\triangle}EF$-hand mutant also showed significant changes in current amplitudes and potentiation time. Our findings suggest that there is a constitutive inhibition of EF-hand and binding of CaM C-lobe on the channel in low calcium concentration. At higher calcium concentration, calcium ions occupy the N-lobe as well as the EF-hand domain, allowing the two to compete to bind to the channel.

• Molecules and Cells
• /
• 제44권1호
• /
• pp.13-25
• /
• 2021

Apoptosis and compensatory proliferation, two intertwined cellular processes essential for both development and adult homeostasis, are often initiated by the mis-regulation of centrosomal proteins, damaged DNA, and defects in mitosis. Fly Anastral spindle 3 (Ana3) is a member of the pericentriolar matrix proteins and known as a key component of centriolar cohesion and basal body formation. We report here that ana3m19 is a suppressor of lethality induced by the overexpression of Sol narae (Sona), a metalloprotease in a disintegrin and metalloprotease with thrombospondin motif (ADAMTS) family. ana3m19 has a nonsense mutation that truncates the highly conserved carboxyl terminal region containing multiple Armadillo repeats. Lethality induced by Sona overexpression was completely rescued by knockdown of Ana3, and the small and malformed wing and hinge phenotype induced by the knockdown of Ana3 was also normalized by Sona overexpression, establishing a mutually positive genetic interaction between ana3 and sona. p35 inhibited apoptosis and rescued the small wing and hinge phenotype induced by knockdown of ana3. Furthermore, overexpression of Ana3 increased the survival rate of irradiated flies and reduced the number of dying cells, demonstrating that Ana3 actively promotes cell survival. Knockdown of Ana3 decreased the levels of both intra- and extracellular Sona in wing discs, while overexpression of Ana3 in S2 cells dramatically increased the levels of both cytoplasmic and exosomal Sona due to the stabilization of Sona in the lysosomal degradation pathway. We propose that one of the main functions of Ana3 is to stabilize Sona for cell survival and proliferation.

• 한국항만경제학회지
• /
• 제38권2호
• /
• pp.69-80
• /
• 2022

우리나라에서 화물 수송분담율을 가장 많이 차지하는 것이 도로운송이다. 도로 컨테이너 운송은 컨테이너 운송의 출발점이자 완결점일 뿐만 아니라 매우 복잡하고 종사자 또한 많다. 육상물류에서 핵심 역할을 하고 있는 컨테이너 운전기사의 본업은 컨테이너를 항만 및 터미널에서부터 목적지까지 운송해 주는 것이다. 그러나 운송과정에서 컨테이너의 청소 및 데미지 컨테이너의 관리 등 본업외에도 여러 가지 컨테이너 관리의무를 묵시적으로 전가받고 있는 실정이다. 그러나 동 사안에 대해서는 그동안 거의 연구가 되고 있지 않은 상황이다. 본 연구는 컨테이너 운전기사의 근로환경에 대해 파악하고, 컨테이너 관리 작업에 대한 실태와 현황을 실증적으로 분석했다. 나아가 선사, 터미널, 화주별로 컨테이너 관리 요구 정도의 차이가 있는지 분석하는 한편 조사지역별 요구사항의 차이를 분석하고, 컨테이너 관리 전가문제에 대한 개선방안을 제시하였다.

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
• /
• pp.114.1-114
• /
• 2003

A new isolate of rod-shaped virus was identified from grafted cactus, Gymnocalycium mihanovichii grafted onto Hylocereus trigonus, in Korea. The virus proved to be a new Tobamovirus and called previously as Tobamovirus-Ca for which we suggest the name Cactus mild mottle virus(CMMoV), because it produced systemic mild mosaic symptoms on its original host. CMMoV is distantly related to known species of the genus Tobamovirus on the basis of host range, serological and sequence analyses. Western blot analysis showed that CMMoV is serologically unrelated to Summons' Opuntia virus which is the only known species of the genus found in cactus plants. The 3'-terminal 2,910 nucleotides have been sequenced for the virus. The coat protein (CP) and movement protein (MP) genes encode 161 and 306 amino acids residues, respectively. The nucleotide and amino acid sequences of the CP were 39.6 ％ to 49.2 ％ and 26.4 ％ to 40.3 ％ identical to other tobamoviruses, respectively. The MP and 3' noncoding region shared 16.3 ％ to 23.3 ％ and 44.6 ％ to 63.4 ％ identities, respectively, with the members of the genus. Phylogenetic tree analysis of the CP gene revealed that CMMoV clusters with members of subgroup I of Tobamovirus. CMMoV particles contained genomic RNA along with two subgenomic RNAs, and this characteristics is common in the members of the subgroup II. This is the first information of sequence and comparative analysis of a Tobamovirus that infects cactus.

• BMB Reports
• /
• 제56권2호
• /
• pp.78-83
• /
• 2023

Chronic myeloid leukemia (CML) has a markedly improved prognosis with the use of breakpoint cluster region-abelson 1 (BCR-ABL1) tyrosine kinase inhibitors (BCR-ABL1 TKIs). However, approximately 40% of patients are resistant or intolerant to BCR-ABL1 TKIs. Hypoxia-inducible factor 1α (HIF-1α) is a hypoxia response factor that has been reported to be highly expressed in CML patients, making it a therapeutic target for BCR-ABL1 TKI-sensitive CML and BCR-ABL1 TKI-resistant CML. In this study, we examined whether HIF-1α inhibitors induce cell death in CML cells and BCR-ABL1 TKI-resistant CML cells. We found that echinomycin and PX-478 induced cell death in BCR-ABL1 TKIs sensitive and resistant CML cells at similar concentrations while the cell sensitivity was not affected with imatinib or dasatinib in BCR-ABL1 TKIs resistant CML cells. In addition, echinomycin and PX-478 inhibited the c-Jun N-terminal kinase (JNK), Akt, and extracellular-regulated protein kinase 1/2 (ERK1/2) activation via suppression of BCR-ABL1 and Met expression in BCR-ABL1 sensitive and resistant CML cells. Moreover, treatment with HIF-1α siRNA induced cell death by inhibiting BCR-ABL1 and Met expression and activation of JNK, Akt, and ERK1/2 in BCR-ABL1 TKIs sensitive and resistant CML cells. These results indicated that HIF-1α regulates BCR-ABL and Met expression and is involved in cell survival in CML cells, suggesting that HIF-1α inhibitors induce cell death in BCR-ABL1 TKIs sensitive and resistant CML cells and therefore HIF-1α inhibitors are potential candidates for CML treatment.

• BMB Reports
• /
• 제40권2호
• /
• pp.247-260
• /
• 2007

Cinnamate 4-hydroxylase (C4H) is a key enzyme of phenylpropanoid pathway, which synthesizes numerous secondary metabolites to participate in development and adaption. Two C4H isoforms, the 2192-bp BnC4H-1 and 2108-bp BnC4H-2, were cloned from oilseed rape (Brassica napus). They both have two introns and a 1518-bp open reading frame encoding a 505-amino-acid polypeptide. BnC4H-1 is 57.73 kDa with an isoelectric point of 9.11, while 57.75 kDa and 9.13 for BnC4H-2. They share only 80.6% identities on nucleotide level but 96.6% identities and 98.4% positives on protein level. Showing highest homologies to Arabidopsis thaliana C4H, they possess a conserved p450 domain and all P450-featured motifs, and are identical to typical C4Hs at substrate-recognition sites and active site residues. They are most probably associated with endoplasmic reticulum by one or both of the N- and C-terminal transmembrane helices. Phosphorylation may be a necessary post-translational modification. Their secondary structures are dominated by alpha helices and random coils. Most helices locate in the central region, while extended strands mainly distribute before and after this region. Southern blot indicated about 9 or more C4H paralogs in B. napus. In hypocotyl, cotyledon, stem, flower, bud, young- and middle-stage seed, they are co-dominantly expressed. In root and old seed, BnC4H-2 is dominant over BnC4H-1, with a reverse trend in leaf and pericarp. Paralogous C4H numbers in Brassicaceae genomes and possible roles of conserved motifs in 5' UTR and the 2nd intron are discussed.

• 생명과학회지
• /
• 제19권2호
• /
• pp.284-288
• /
• 2009

선천성 면역은 숙주의 물리적 방어벽을 뚫고 침입하는 감염성 질병 원인균에 대항하는 첫 번째 방어로서 아주 중요한 역할을 한다. Mannose-binding lectin (MBL 또는 mannan-binding protein, MBP)은 혈청 내에 존재하는 면역성 단백질로서 감염 후 즉시 유발되는 acute phase response의 특정 단백질이다. MBL 단백질은 세균, 바이러스, 곰팡이, 기생충 등의 탄수화합물 구조에 결합하여 식균 작용을 돕거나 보체경로를 활성화 시킨다. MBL 단백질은 C-말단이 탄수화물을 인식하는 도메인이며, 연결 목 부위와 콜라겐 부위로 구성되어 있다. 혈청 내의 MBL 농도가 낮으면 높은 빈도로 면역결핍현상이 관찰된다고 알려져 있다. MBL 단백질의 기능과 유전에 대해 많은 연구가 되어져 왔으나 아직 MBL 단백질 복합체 등에 대한 연구는 많이 이루어져 있지 않다. 따라서 MBL 연구에 필수적인 MBL cDNA 제조와 재조합 단백질의 합성, 그리고 재조합 단백질을 항원으로 사용하여 polyclonal antibody를 생산한 연구 결과를 보고하는 바이다. 본 연구결과로 획득한 MBL cDNA, 재조합 단백질과 anti-MBL 항체는 앞으로의 MBL 연구에 절대적으로 필요한 도구가 될 것으로 생각된다.