• Molecules and Cells
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• 제46권6호
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• pp.374-386
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• 2023

Thermal stress induces dynamic changes in nuclear proteins and relevant physiology as a part of the heat shock response (HSR). However, how the nuclear HSR is fine-tuned for cellular homeostasis remains elusive. Here, we show that mitochondrial activity plays an important role in nuclear proteostasis and genome stability through two distinct HSR pathways. Mitochondrial ribosomal protein (MRP) depletion enhanced the nucleolar granule formation of HSP70 and ubiquitin during HSR while facilitating the recovery of damaged nuclear proteins and impaired nucleocytoplasmic transport. Treatment of the mitochondrial proton gradient uncoupler masked MRP-depletion effects, implicating oxidative phosphorylation in these nuclear HSRs. On the other hand, MRP depletion and a reactive oxygen species (ROS) scavenger non-additively decreased mitochondrial ROS generation during HSR, thereby protecting the nuclear genome from DNA damage. These results suggest that suboptimal mitochondrial activity sustains nuclear homeostasis under cellular stress, providing plausible evidence for optimal endosymbiotic evolution via mitochondria-to-nuclear communication.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 추계국제학술대회
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• pp.55-55
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• 2020

뫼제비꽃(Viola selkirkii)의 엽록체 DNA 염기서열을 차세대염기서열분석법(NGS)을 이용하여 분석하였다. 재료는 강원도 화천군 일산과 제주도 한라산의 2개체를 사용하였다. 분석결과, 염기서열의 길이는 일산의 뫼제비꽃이 156,774 bp (GC content: 36.30%), 한라산의 뫼제비꽃이 157,451 bp(GC content: 36.30%)로 한라산 개체가 길게 분석되었다. 구간별로 LSC(Large single copy)지역은 한라산 개체(85,950 bp)가 일산 개체(85,930 bp)보다 20 bp 길었으며, SSC(Small single copy)지역은 한라산 개체(17,261 bp)보다 일산 개체가 17,982 bp로 길게 분석되었다. IR(Inverted repeat)지역은 한라산 개체가 27,120 bp로 일산 개체(26,431 bp)보다 길게 분석되었다. 이러한 염기서열 길이의 차이는 종내 개체 간 빈번하게 발생하는 현상으로 IGS와 intron 구간에서 확인 된 단순반복서열의 일부 누락과 IR지역 내의 수축과 확장에 의한 것으로 판단된다. 뫼제비꽃 2개체의 엽록체 게놈을 구성하는 유전자 수는 총 111개로 동일하였으며, protein coding gene 77개, tRNA(transfer RNA) gene 30개, 그리고 rRNA (ribosomal RNA) gene 4개로 구성되어 있었다. 이는 기 발표된 엽록체 DNA 전체 염기서열이 밝혀진 제비꽃속 (Viola) 종류들과 동일한 결과이다.

• ALGAE
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• 제39권1호
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• pp.17-30
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• 2024

Red macroalga Pyropia yezoensis is a high valuable cultivated marine crop. Its acclimation to cold stress is especially important for long cultivation period across winter in coasts of warm temperate zone in East Asia. In this study, the response of P. yezoensis thalli to low temperature was analyzed on physiology and transcriptome level, to explore its acclimation mechanism to cold stress. The results showed that the practical photosynthesis activity (indicated by Φ_PSII and qP) was depressed and pigment allophycocyanin content was decreased during the cold stress of 48 h. However, the F_v/F_m and non-photochemical quenching increased significantly after 24 h, and the average growth rate of thalli also rebounded from 24 to 48 h, indicating a certain extent of acclimation to cold stress. On transcriptionally, the low temperature promoted the expression of differentially expressed genes (DEGs) related to carbohydrate metabolism and energy metabolism, while genes related to photosynthetic system were depressed. The increased expression of DEGs involved in ribosomal biogenesis and lipid metabolism which could accelerate protein synthesis and enhance the degree of fatty acid unsaturation, might help P. yezoensis thallus cells to cope with cold stress. Further co-expression network analysis revealed differential expression trends along with stress time, and corresponding hub genes play important roles in the systemic acquired acclimation to cold stress. This study provides basic mechanisms of P. yezoensis acclimation to cold temperature and may aid in exploration of functional genes for genetic breeding of economic macroalgae.

• 한국군사과학기술학회지
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• 제27권2호
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• pp.304-310
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• 2024

Ricin is a highly toxic protein which is produced in the seeds of the castor oil plant. Ricin toxin A chain has ribosomal RNA N-glycosylase activity that irreversibly hydrolyses the N-glycosidic bond of the adenine residue at position 4324 within the 28S rRNA. In this study, we developed non-toxic recombinant ricin vaccine(R51) in E. coli expression system, and evaluated efficacy of the R51 according to adjuvants. When the R51 was administered using aluminum hydroxide as an adjuvant, the vaccine efficacy was higher than that of TLR agonists or aluminum phosphate. Because it is time-consuming to administer the vaccine three times at three-week intervals, we investigated the survival rate and antibody titer of mice according to the change of time interval of vaccination. Interestingly, there was no difference in survival rate and antibody titer when R51 was administered at 0, 1, and 3 weeks or 0, 2, and 4 weeks compared to when administered at 0, 3, and 6 weeks. Therefore, the developed R51 vaccine is promising to protect soldiers from Ricin attack.

• 한국패류학회지
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• 제29권4호
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• pp.335-341
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• 2013

본 연구는 우리나라의 주요양식 품종인 북방전복을 대상으로 지금까지 전복류에서는 사용되지 않았던 mtDNA의 protein coding 영역 ND2, ND5, ND4, ND4L, ND6, ND1의 6개영역과 protein noncoding 영역인 12SrRNA(ribosomal RNA) 을 포함해 총 7개 영역을 이용하여 각 영역의 유전적 변이성 및 개체간 유전적 유연관계 등을 분석하여 각 영역별 특성을 파악하고 이러한 특성을 고려하여 유전학적 분석에 적합한 분자유전마커를 개발하였다. 유전적 변이성은 ND4 영역 (Haplotype diversity = 1.000, Nucleotide diversity = 0.010823) 이 가장 높게 나타났으며, 개체간의 유전적 차이는 ND2 및 ND1 영역이 각각 90% 및 87%로 유의적으로 명확히 구분할 수 있었다. 따라서 유전적 변이성이 가장 높은 ND4 영역과 영역내의 클러스터 간의 유전적 차이가 명확한 ND2 및 ND1 영역을 복합적으로 활용할 경우 북방 전복의 집단유전학 및 계통분류학 분석에 유용한 분자유전마커로 사용할 수 있을 것이라 생각된다.

• Parasites, Hosts and Diseases
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• 제51권6호
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• pp.719-726
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• 2013

Mitochondrial genomes have been extensively studied for phylogenetic purposes and to investigate intra- and interspecific genetic variations. In recent years, numerous groups have undertaken sequencing of platyhelminth mitochondrial genomes. Haplorchis taichui (family Heterophyidae) is a trematode that infects humans and animals mainly in Asia, including the Mekong River basin. We sequenced and determined the organization of the complete mitochondrial genome of H. taichui. The mitochondrial genome is 15,130 bp long, containing 12 protein-coding genes, 2 ribosomal RNAs (rRNAs, a small and a large subunit), and 22 transfer RNAs (tRNAs). Like other trematodes, it does not encode the atp8 gene. All genes are transcribed from the same strand. The ATG initiation codon is used for 9 protein-coding genes, and GTG for the remaining 3 (nad1, nad4, and nad5). The mitochondrial genome of H. taichui has a single long non-coding region between trnE and trnG. H. taichui has evolved as being more closely related to Opisthorchiidae than other trematode groups with maximal support in the phylogenetic analysis. Our results could provide a resource for the comparative mitochondrial genome analysis of trematodes, and may yield genetic markers for molecular epidemiological investigations into intestinal flukes.

• Asian-Australasian Journal of Animal Sciences
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• 제14권6호
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• pp.880-884
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• 2001

Rumen microbiology research has undergone several evolutionary steps: the isolation and nutritional characterization of readily cultivated microbes; followed by the cloning and sequence analysis of individual genes relevant to key digestive processes; through to the use of small subunit ribosomal RNA (SSU rRNA) sequences for a cultivation-independent examination of microbial diversity. Our knowledge of rumen microbiology has expanded as a result, but the translation of this information into productive alterations of ruminal function has been rather limited. For instance, the cloning and characterization of cellulase genes in Escherichia coli has yielded some valuable information about this complex enzyme system in ruminal bacteria. SSU rRNA analyses have also confirmed that a considerable amount of the microbial diversity in the rumen is not represented in existing culture collections. However, we still have little idea of whether the key, and potentially rate-limiting, gene products and (or) microbial interactions have been identified. Technologies allowing high throughput nucleotide and protein sequence analysis have led to the emergence of two new fields of investigation, genomics and proteomics. Both disciplines can be further subdivided into functional and comparative lines of investigation. The massive accumulation of microbial DNA and protein sequence data, including complete genome sequences, is revolutionizing the way we examine microbial physiology and diversity. We describe here some examples of our use of genomics- and proteomics-based methods, to analyze the cellulase system of Ruminococcus flavefaciens FD-1 and explore the genome of Ruminococcus albus 8. At Illinois, we are using bacterial artificial chromosome (BAC) vectors to create libraries containing large (>75 kbases), contiguous segments of DNA from R. flavefaciens FD-1. Considering that every bacterium is not a candidate for whole genome sequencing, BAC libraries offer an attractive, alternative method to perform physical and functional analyses of a bacterium's genome. Our first plan is to use these BAC clones to determine whether or not cellulases and accessory genes in R. flavefaciens exist in clusters of orthologous genes (COGs). Proteomics is also being used to complement the BAC library/DNA sequencing approach. Proteins differentially expressed in response to carbon source are being identified by 2-D SDS-PAGE, followed by in-gel-digests and peptide mass mapping by MALDI-TOF Mass Spectrometry, as well as peptide sequencing by Edman degradation. At Ohio State, we have used a combination of functional proteomics, mutational analysis and differential display RT-PCR to obtain evidence suggesting that in addition to a cellulosome-like mechanism, R. albus 8 possesses other mechanisms for adhesion to plant surfaces. Genome walking on either side of these differentially expressed transcripts has also resulted in two interesting observations: i) a relatively large number of genes with no matches in the current databases and; ii) the identification of genes with a high level of sequence identity to those identified, until now, in the archaebacteria. Genomics and proteomics will also accelerate our understanding of microbial interactions, and allow a greater degree of in situ analyses in the future. The challenge is to utilize genomics and proteomics to improve our fundamental understanding of microbial physiology, diversity and ecology, and overcome constraints to ruminal function.

• 한국어류학회지
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• 제18권2호
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• pp.65-77
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• 2006

우리나라 주요 담수 어종인 미꾸라지(Misgurnus mizolepis)를 실험 모델로 이용하여 실험적으로 설정한 고온 노출($32^{\circ}C$)에 특이적으로 반응하는 유전자들을 cDNA microarray 분석을 통해 탐색하였다. 미꾸라지 간조직 expressed sequence tag (EST) 데이터베이스 분석을 통해 1,124개의 unigene들을 선발하여 제작한 cDNA microarray을 이용하여 $23^{\circ}C$ 및 $32^{\circ}C$에 4주간 노출된 실험어의 간(liver)조직의 전사 발현 양상을 3반복 분석하였다. 다양한 유전자군이 $32^{\circ}C$ 고온 노출에 전사 발현의 증감 또는 감소 양상을 보였으며 $23^{\circ}C$에 비해 $32^{\circ}C$군에서 2배 이상의 발현 증가를 보인 클론들은 총 93종류로서 에너지 대사, 단백질 대사, 면역/항산화 기능, 세포골격 및 구조, 물질수송 및 세포 신호전달등에 관여하는 단백질들을 암호화하는 유전자들이었고 최대 15배 이상의 전사발현이 관찰되었다. 반면 고온 노출군에서 유의적인 발현 감소(50% 이하)를 보인 유전자들(n=85) 역시 탐색되어 상기 단백질 분류군외에 vitellogenin 전구체들 및 리보좀 단백질류에서 특이적인 전사활성의 저하가 관찰되었고, vitellogenin 유전자에서 가장 많은 mRNA 수준의 감소가 관찰되었다.

• 한국임상수의학회지
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• 제28권5호
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• pp.516-518
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• 2011

한국 황새복원센터에서 사육중인 황새의 정상 장내 세균총을 검사하기 위한 조사를 실시하였다. 44마리의 건강한 황새의 총배설강으로부터 44개의 분변시료를 채취하여 호기 및 혐기조건에서 배양하였다. 배양된 미생물의 16Sribosomal RNA 및 heat shock protein 60 유전자의 염기서열을 조사하였다. 호기조건 하에서는 Enterococcus faecalis, Escherichia coli, Bacillus spp., Enterococcus avium, Enterococcus gallinarum, Pseudomonas spp., Alcaligenes spp., Enterobacter spp., Corynebacterium spp., Proteus mirabilis가 동정되었다. 혐기조건 하에서는 E. coli, Clostridium tertium, En. faecalis, P. mirabilis가 동정되었다. 모든 시료에서 E. coli, En. faecalis 혹은 두 가지 모두가 분리되었다. 이러한 정상 황새의 장내 미생물 정보는 향후 분변 배양 결과의 해석에 도움이 될 것으로 판단된다.