Histone Lysine Methylation

곽상준,Kwak, Sahng-June Korean Society of Life Science 2007 생명과학회지 Vol.17 No.3

Our genome exists in the form of chromatin, and its structural organization should be precisely regulated with an appropriate dynamic nature for life. The basic unit of chromatin is a nucleosome, which consists of a histone octamer. These nucleosomal histones are subject to various covalent modifications, one of which is methylation on certain lysine residues. Recent studies in histone biology identified many histone Iysine methyltransferases (HKMTs) responsible for respective lysine residues and uncovered various kinds of involved chromatin associating proteins and many related epigenetic phenotypes. With the aid of highly precise experimental tools, multi-disciplinary approaches have widened our understanding of how lysine methylation functions in diverse epigenetic processes though detailed mechanisms remain elusive. Still being considered as a relatively more stable mark than other modifications, the recent discovery of lysine demethylases will confer more flexibility on epigenetic memory transmitted through histone lysine methylation. In this review, advances that have been recently observed in epigenetic phenotypes related with histone lysine methylation and the enzymes for depositing and removing the methyl mark are provided.

Current Progress in Generation of Genetically Modified Mice

송기덕,조병욱,Song, Ki-Duk,Cho, Byung-Wook Korean Society of Life Science 2007 생명과학회지 Vol.17 No.4

Manipulation of the mouse genome by activating or inactivating the gene has contributed to the understanding of the function of the gene in the subset of cells during embryonic development or postnatal period of life. Most of all, gene targeting, which largely depends on the availability of mouse embryonic stem (ES) cells, is the milestone of development of animal models for human disease. Recombinase-mediated genome modification (Cre-LoxP and Flp-Frt etc) and the ligand-dependent regulation system, more accurate and elaborate manipulation tools, have been successfully developed and applied to dissect the mechanisms governing complex biological processes and to understand the role of protein in temporal-and spatial aspects of development. As technologies concerning refined manipulation of mouse genome are developed, they are expected to open new opportunities to better understand the diverse in vivo functions of genes.

Genetic Variation and Population Structure of Alder (Alnus hirsuta : Betulaceae) in Korea

Park, Joo-Soo,Huh, Man-Kyu Korean Society of Life Science 2000 Journal of Life Science Vol.10 No.1

Variation at 25 allozyme loci in Korean wateralder (Alnus hirsuta Rupr.) from nine distinct populations was measured to estimate the amount and pattern of genetic diversity and population structure. The mean genetic diversity within population was 0.166. Korean alder populations have slightly high levels of genetic diversity compared to those present in associated temperature-zone species and two Canadian alder species. Among population s genetic differentiation accounted for an significant 9% of the total variation. High gene flow(Nm=2.63) was observed. Analysis of fixation indices, calculated for all polymorphic loci in each population, showed a substantial deficiency of heterozygotes relative to Hardy-Weinberg expectations. The mean GST value A. hirsuta in Korea (GST = 0.087) is similar to those of A. rogosa in Canada (GST = 0.052). These low values of GST in two countries. reflecting little spatial genetic differentiation, may indicate extensive gene flow (via pollen and/or seeds) and/or recent colonization. These factors reduce the effect of geographic isolation of breeding and the chance for genetic divergence. A pattern of increasing is observed with increasing rainfall per year. Regression analysis indicates that 54% (F = 4.67) of the variability observed can be explained by this relationship.

Expression analysis of RBMY1, CDY1, and VCY2 genes in Korean male infertility

허재원,김우영,김대수,하홍석,박남철,최욱환,남기만,최진,김희수,Huh, Jae-Won,Kim, Woo-Young,Kim, Dae-Soo,Ha, Hong-Seok,Park, Nam-Chul,Choi, Ook-Hwan,Nam, Ki-Man,Choi, Jin,Kim, Heui-Soo Korean Society of Life Science 2007 생명과학회지 Vol.17 No.5

Azoospermia factor(AZFa, b, and c) regions have been focused on their involvement in the spermatogenic process by frequent observation of microdeletion in male infertility. Among the azoospermia factors, RBMY1, CDY1, and VCY2 genes are strongly associated with the male germinal cell differentiation and development in testis. Using RT-PCR approach, expression patterns of RBMY1, CDY1, and VCY2 genes are examined in testicular biopsy specimens from 42 Korean azoospermic patients. No expression of RBMY1, CDY1, and VCY2 genes appeared as 34%, 66%, and 27% of the male infertility, respectively. Patients who had no expression of RBMY1 and VCY2 genes also showed negative expression of the CDY1 gene in their testis tissues. All Sertoli cell-only syndrome patients showed no expression of the CDY1 gene. Taken together, the CDY1 gene expression seems to be necessary factor to complete spermatogenesis in Korean population.

Antifungal Activity of Korean Radish (Raphanus sativaus L) Extracts Against Pathogenic Plant

Won, Hwang-Cher- Korean Society of Life Science 2003 생명과학회지 Vol.13 No.2

A study of the anti-fungal properties in Korean radish was conducted using a variety of purification procedures such as Extrelut column, RP(Reverse Phase) Cl8 Column Chromatography, HPLC etc. to separate anti-fungal substances from Korean radish juices to test them against a common gray mold called Botrytis cenerea. Dialysis tube operation showed that these substances were presumably thermostable compounds with low molecular mass (less than 3.5 kDa). Differences of anti-fungal activities depending upon types of radishes used did not show any noticeable variation. The antifungals were presumably composed of more than 5 compounds. Among these, the most anti-fungal fraction was analyzed by HPLC in which one peak was obtained. Disease-affected plants were inoculated with 10mg of Extrelut fraction and results showed similar anti-fungal activity to pesticides suggesting possible usage of these substances as environmentally friendly antibiotics.

Phylogenetic Relationship among Several Korean Coastal Red Tide Dinoflagellates Based on their rDNA Internal Transcribed Spacer Sequences

Cho, Eun-Seob,Kim, Gi-Yong,Park, Hyung-Sik,Nam, Byung-Hyouk,Lee, Jae-Dong Korean Society of Life Science 2001 Journal of Life Science Vol.11 No.2

The nucleotide sequences of the internal transcribed spacer regions (ITS1 and ITS2) of ribosomal DNA (rDNA), and the 5.85 rRNA gene, have been determined for 13 strains of dinoflagellates in order to analyze the phylo-genetic relationship. The DNA sequences contained considerable variation in the ITS regions, but little in the 5.85 rDNA. In addition, the ITS1 was more variable than the ITS2 in all species examined. The nucleotide length of this region varied from 519 bp to 596 bp depending on the taxa. The investigated taxa were divided into three large groups based on the ITS length, i. e., a group with short ITS region (A. fraterculus and Alexandrium sp.), a with ITS region group (P. micans, P. minimum and P. triestinum) and a with ITS region group (G. impudicum, C. polykrikoides, G. sanguineum, G. catenatum and H. triquetra). The relationship between nucleotide length of ITS1 and that of ITS2 was negative, whereas G+C content and nucleotide length showed positive correlation. In phylogenetic analyses producing NJ trees, the topology was similar cluster and clearly divided the taxa into three groups based on 5.8S rDNA that were similar to those based on morphological characteristics. In particular, G. impudicum was more closely related to G. catenatum than to C. polykrikoides using phylogenetic analysis. From this study, we chew that the length of ITS region contributes to discriminate Korean harmful algal species and ITS analysis is a useful method for resolving the systematic relationships of dinoflagellates.

Molecular phylogeny of moon jellyfish Aurelia aurita Linnaeus collected from Yeosu waters in Korea based on nuclear and mitochondrial DNA sequences

김숙양,조은섭,Kim, Sook-Yang,Cho, Eun-Seob Korean Society of Life Science 2007 생명과학회지 Vol.17 No.3

This study presents the molecular phylogenetic analysis of Korean Aurelia aurita Linnaeus collected from Yeosu in the southern waters of Korea using nuclear ITS1 region and mitochondrial COI gene sequences. The use of oligonucleotide primers F5 (forward) and R5 (reverse) targeted to ITS1 and LCO1490 (forward) and HCO2198 (reverse) targeted to COI amplified 267 bp and 643 bp fragments, respectively. The shortest genetic distance towards the ITS1 region is estimated at 0.023 when comparing Korean A. aurita to Aurelia sp. collected from California, USA. In particular, Korean and American/Swedish A. aurita were located far away in terms of genetic distance, ranging from 0.393 to 0.395. On the other hand, the genetic distance between Korean and English/Turkish/Swedish/American A. aurita regarding the mitochondrial DNA COI gene ranged from 0.201 to 0.205. However, a sister-ship with Korean and American A. aurita showed an extremely high bootstrap value (100%). The predicted secondary RNA structure of the mitochondrial DNA COI gene showed many different folding structures with a similar energy between Korean and American A. aurita. These results suggest that ITS1 and the mitochondrial DNA COI gene could be used as genetic markers for identification of the biogeographic populations.

Population Genetic Structure of Japanese Anchovy (Engraulis japonicus) in Korean waters Based on Mitochondrial 12S Ribosomal RNA Gene Sequences

김진영,조은섭,김우진,Kim Jin Yeong,Cho Eun Seob,Kim Woo Jin Korean Society of Life Science 2004 생명과학회지 Vol.14 No.6

We used portions of mitochondrial 125 ribosomal RNA gene sequences (339 bp) to investigate the phylogenetic and population genetic characteristics of the Japanese anchovy, Engraulis japonicus, in Korean waters. A total of 35 mtDNA haplotypes were obtained from the samples collected in 3 locations (the southern area of the Yellow Sea, the western coast of Jejudo, and the eastern area of the South Sea) in Korean waters. One haplotype, AN8T103, obtained from the southern area of the Yellow Sea, was formed according to an independent phylogenetic individual in the PAUP analysis, which was separated from the others by a $0.2-4.1\%$ sequence divergence. This distinct haplotype appeared to be one that was carried by immigrants from another study area, but further study is necessary. Genetic divergence, except for AN8T103, was moderate to substantial $(0.2-3.8\%)$ and nucleotide diversity within populations was 0.015 for Yellow Sea, 0.013 for Jejudo, and 0.D15 for South Sea, respectively. The female gene flow was substantial or high (Nm=25.5-36.4), and the genetic distances between regions were not statistically significant $(P>0.01)$. These results indicated that the Japanese anchovy populations occurring in Korean waters were consisted of individuals randomly dispersed over geographic areas.

Adenoviral-Mediated Ref-1 Overexpression Potentiates NO Production in Bradykinin-Stimulated Endothelial Cells

송주동,김강미,이상권,김종민,박영철,Song, Ju-Dong,Kim, Kang-Mi,Lee, Sang-Kwon,Kim, Jong-Min,Park, Young-Chul Korean Society of Life Science 2007 생명과학회지 Vol.17 No.7

The dual-function protein redox factor-1 (Ref-1) is essential for base excision repair of oxidatively damaged DNA and also governs the activation of many redox-sensitive transcription factors. We examined the role of Ref-1 in regulation of nitric oxide (NO) synthesis employing adenoviral-mediatedoverexpression of Ref-1 in bradykinin-stimulated endothelial cells. Intracellular NO was detected with the NO-sensitive fluorophore DAF-2. Overexpression of Ref-1 potentiates bradykinin-stimulated NO production in endothelial cells. And, cells ifected with AdRef-1 showed higher fluorescence intensity compared with uninfected or AdD1312-infected cells. In parallel with this, over expression of Ref-1 also stimulated endothelial NO synthase (eNOS) enzyme activity, compared with unifected or AdD1312-infected cells, in bradykinin-stimulated cells as well as in unstimulated cells. These results suggest that Ref-1 implicates in endothelium-dependent vasorelaxation resulting from NO production in vascular system.

The Study of Trnascriptional Regulated Gene, $hrp^{2+}$, in Yeast

Choi, In-Soon Korean Society of Life Science 2001 Journal of Life Science Vol.11 No.2

This study was designed to clone the SNF2/SW12 helicase-related genes from the fission yeast Schizosaccha-romyces pombe and thereafter to elucidate the common functions of the proteins in this family. The $hrp^{2+}$gene was cloned by polymerase chain reaction amplification using degenerative primers from conserved SNF2 motifs within the ERCC6 gene, which encodes a protein involved in DNA excision repair. Like other SNF2/SW12 family proteins, the deduced amino acid sequence of Hrp2 contains DNA-dependent ATPase/7 helicase domains as well as the chromodomain and the DNA binding domain. This configuration is similar to that of mCHD1 (mouse chromo-ATPase/helicase-DNA-dinding protein 1), suggesting that Hrp2 is a S. pombe homolog of mCHD1, which is thought to function in altering the chromatin structure to control the gene expression. To characterize the function of Hrp2, 4 Uracil-Hrp2 fusion protein, it was purified near homogeneity by affinity chromatography on $Ni^{2+}$-NTA agarose, DEAE-Sepharose ion exchange arid Sephacryl S-200 gel filtration chromatographies. The purified fusion protein exhibited DNA-dependent ATPase activity, which was stimulated by both double-stranded and single-stranded DNA. To determine the steady-state level of $hrp^{2+}$ transcripts during growth, cells were cultured in medium and collected at every 2hr to prepare total RNAs. The northern blot analysis showed that the level of $hrp^{2+}$ transcripts reached its maximum before the cells entered the exponential growth phase and then decreased gradually, This result implies that Hrp2 may be required at early stages of cell growth.h.