• Title/Summary/Keyword: noncoding DNA

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Identification of a New 5'-Noncoding Exon Region and Promoter Activity in Human N-Acetylglucosaminyltransferase III Gene

  • Kang, Bong-Seok;Kim, Yeon-Jeong;Shim, Jae-Kyoung;Song, Eun-Young;Park, Young-Guk;Lee, Young-Choon;Nam, Kyung-Soo;Kim, June-Ki;Lee, Tae-Kyun;Chung, Tae-Wha;Kim, Cheorl-Ho
    • BMB Reports
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    • v.31 no.6
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    • pp.578-584
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    • 1998
  • In a previous paper (Kim et al., 1996a), the immediate 5' -flanking region and coding region of the human UDP-N -acetylglucosamine:-D-mannoside-1,4-Nacetylglucosaminyltransferase III (N-acetylglucosaminyitransferase- III; GnT-III) gene was reported, isolated and analyzed. Herein, we report on amplification of a new 5' -noncoding region of the GnT-III mRNA by single-strand ligation to single-stranded cDNA-PCR (5' -RACE PCR) using poly(A)+ RNA isolated from human fetal liver cells. A cDNA clone was obtained with 5' sequences (96 bp) that diverged seven nucleotides upstream from the ATG (+1) start codon. A concensus splice junction sequence, TCTCCCGCAG, was found immediately 5' to the position where the sequences of the cDNA diverged. The result suggested the presence of an intron in the 5' -noncoding region and that the cDNA was an incompletely reversetranscribed cDNA product derived from an mRNA containing a new noncoding exon. When mRNA expression of GnT-III in various human tissues and cancer cell lines was examined, Northern blot analysis indicated high expression levels of GnT-III in human fetal kidney and brain tissues, as well as for a number of leukemia and lymphoma cancer cell lines. Promoter activities of the 5' -flanking regions of exon 1 and the new noncoding region were measured in a human hepatoma cell line, HepG2, by luciferase assays. The 5'-flanking region of exon 1 was the most active, whilst that of exon 2 was inactive.

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DNA Information Hiding Method for DNA Data Storage (DNA 데이터 저장을 위한 DNA 정보 은닉 기법)

  • Lee, Suk-Hwan;Kwon, Ki-Ryong
    • Journal of the Institute of Electronics and Information Engineers
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    • v.51 no.10
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    • pp.118-127
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    • 2014
  • DNA data storage refers to any technique for storing massive digital data in base sequence of DNA and has been recognized as the future storage medium recently. This paper presents an information hiding method for DNA data storage that the massive data is hidden in non-coding strand based on DNA steganography. Our method maps the encrypted data to the data base sequence using the numerical mapping table and then hides it in the non-coding strand using the key that consists of the seed and sector length. Therefore, our method can preserve the protein, extract the hidden data without the knowledge of host DNA sequence, and detect the position of mutation error. Experimental results verify that our method has more high data capacity than conventional methods and also detects the positions of mutation errors by the parity bases.

Mitochondrial noncoding RNA transport

  • Kim, Kyoung Mi;Noh, Ji Heon;Abdelmohsen, Kotb;Gorospe, Myriam
    • BMB Reports
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    • v.50 no.4
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    • pp.164-174
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    • 2017
  • Mitochondria are cytosolic organelles essential for generating energy and maintaining cell homeostasis. Despite their critical function, the handful of proteins expressed by the mitochondrial genome is insufficient to maintain mitochondrial structure or activity. Accordingly, mitochondrial metabolism is fully dependent on factors encoded by the nuclear DNA, including many proteins synthesized in the cytosol and imported into mitochondria via established mechanisms. However, there is growing evidence that mammalian mitochondria can also import cytosolic noncoding RNA via poorly understood processes. Here, we summarize our knowledge of mitochondrial RNA, discuss recent progress in understanding the molecular mechanisms and functional impact of RNA import into mitochondria, and identify rising challenges and opportunities in this rapidly evolving field.

Cloning and Nucleotide Sequence of a cDNA Encoding the Rat Triosephosphate Isomerase

  • Lee, Kyunglim;Ryu, Jiwon
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.497-501
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    • 1996
  • A gene coding for triosephosphate isomerase (TPI) from a rat skeletal muscle cDNA library was cloned and its nucleotide sequence was determined. The 1, 348-bp cDNA clone contains 24 bp $5^I$ noncoding region, the entire 750 bp coding region corresponding to a protein of 249 amino acids, $547bp 3^I$ noncoding region and part of a poly(A) tail. It also contains a polyadenylation signal, AATAAA, starting from 17 bp upstream of the poly(A) tail. The calculated molecular weight of rat TPI is 27.8 kDa and the net charge is +4. The deduced amino acid sequence from rat TPI CDNA sequence has 93% and 94% homology with that of mouse and human clones, respectively. The amino acids at the residue of Asn12, Lys14, His96, Glu 166, His96, His101, Ala177, Tyr165, Glu13O, Tyr2O9, and Ser212 in catalytic site are completely identical, confirming that the functional residues in TPI proteins are highly conserved throughout evolution. The most profound characteristic of rat TPI enzyme, compared with other TPIs, is that there are five cysteine substitutions at the residue of 21, 27, 159, 195 and 204. A Glu123 instead of Gly was found in rabbit, rhesus, mouse and human sequences. Through the method of RT-PCR, the mRNA transcription level of TPI gene was found to be different among various tissues and was highest in muscle.

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Phylogenetic Analysis by RFLP and Sequencing of Mitochondrial DNA in a Korean Population

  • Lee, Jin-Young;Kim, Heui-Soo;Ha, Bae-Jin;Park, Yeong-Hong
    • Archives of Pharmacal Research
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    • v.29 no.1
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    • pp.88-95
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    • 2006
  • Analysis of molecular nature of mitochondrial DNA (mtDNA) could be powerful marker for anthropological studies of modern populations. While population genetic studies on mtDNA have been reported for several ethnic groups, no such study has been documented for the Korean population. We surveyed mtDNA polymorphisms in the HVS I of noncoding D-loop region and its upstream region from 430 unrelated healthy Korean population by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing analysis. PCR product with 2,790 bp spanning the specific mtDNA region (mt13715-16504) was subjected to RFLP analysis using 6 restriction enzyme (Hinf I, Hae III, Alu I, Dde I, Mbo I, Rsa I). On the PAUP analysis of PCR-RFLP results, 38 mtDNA haplotypes (Hap 1-38) were detected in the Korean populations, which were classified into 11 haplogroups (Grp 1-11) of related haplotypes encompassing all 38 haplotypes. In comparison of sequencing data with Anderson's reference sequence, the transition type was more prevalent than the transversion type. Insertions or deletions were not found. In addition, three of the polymorphic sites (A16240C, A16351G, G16384A) in HVS-I region are determined newly. The polymorphic sites were distributed randomly in the region, though the frequency at each site was variable. Thus, this research might be required for the genealogical study of Orientals.

Molecular Characterization and Expression Analysis of S6K1 in Cashmere Goats (Capra hircus)

  • Wu, Manlin;Bao, Wenlei;Hao, Xiyan;Zheng, Xu;Wang, Yanfeng;Wang, Zhigang
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.8
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    • pp.1057-1064
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    • 2013
  • p70 ribosomal S6 kinase (p70S6K) can integrate nutrient and growth factor signals to promote cell growth and survival. We report our molecular characterization of the complementary DNA (cDNA) that encodes the goat p70S6K gene 40S ribosomal S6 kinase 1 (S6K1) (GenBank accession GU144017) and its 3' noncoding sequence in Inner Mongolia Cashmere goats (Capra hircus). Goat S6K1 cDNA was 2,272 bp and include an open reading frame (ORF) of 1,578 bp, corresponding to a polypeptide of 525 amino acids, and a 694-residue 3' noncoding sequence with a polyadenylation signal at nucleotides 2,218 to 2,223. The relative abundance of S6K1 mRNA was measured by real-time PCR in 6 tissues, and p70S6K expression was examined by immunohistochemistry in heart and testis. The phosphorylation of p70S6K is regulated by mitogen-activated protein kinase (MAPK) signaling in fetal fibroblasts.

Consecutive Difference Expansion Based Reversible DNA Watermarking (연속적 차분 확장 기반 가역 DNA 워터마킹)

  • Lee, Suk-Hwan;Kwon, Ki-Ryong
    • Journal of the Institute of Electronics and Information Engineers
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    • v.52 no.7
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    • pp.51-62
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    • 2015
  • Of recent interests on high capacity DNA storage, DNA watermarking for DNA copyright protection, and DNA steganography for DNA secret communication are augmented, the reversible DNA watermarking is much needed both to embed the watermark without changing the functionality of organism and to perfectly recover the host DNA sequence. In this paper, we address two ways of DE based reversible DNA watermarking using noncoding DNA sequence. The reversible DNA watermarking should consider the string structure of a DNA sequence, the organism functionality, the perfect recovery, and the high embedding capacity. We convert the string sequence of four characters in noncoding region to the decimal coded values and embed the watermark bit into coded values by two ways; DE based multiple bits embedding (DE-MBE) using pairs of neighbor coded values and consecutive DE-MBE (C-DE-MBE). Two ways process the comparison searching to prevent the false start codon that produces false coding region. Experimental results verified that our ways have more high embedding capacity than conventional methods and produce no false start codon and recover perfectly the host sequence without the reference sequence. Especially C-DE-MBE can embed more high two times than DE-MBE.

Identification of a Novel PGE2 Regulated Gene in SNU1 Gastric Cancer Cell

  • Park, Min-Seon;Kim, Hong-Tae;Min, Byung-Re;Kimm, Ku-Chan;Nam, Myeong-Jin
    • BMB Reports
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    • v.33 no.2
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    • pp.184-187
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    • 2000
  • Prostaglandin $E_2$ ($PGE_2$) plays an important role in the regulation of various gastric functions, and the growth-inhibitory activities on tumor cells are studied in vitro and in vivo. Although the mechanisms have attracted many researchers in the past decade, the molecular mechanisms of cell cycle arrest, or induction of apoptosis by $PGE_2$, is unclear. We investigated the effects of $PGE_2$ on the growth of the human gastric carcinoma cell line SNU1 and genes that are regulated by $PGE_2$ and isolated them using differential display RT-PCR (DD RT-PCR). FACS analysis suggested that SNU1 cells were arrested at the G1 phase by $PGE_2$ treatment. This growth inhibitory effect was in a time- and dose-dependent manner. Treatment of SNU1 cells with $10\;{\mu}g/ml$ $PGE_2$, followed by DD RT-PCR analysis, revealed differently expressed bands patterns from the control. Among the differently expressed clones, we found an unidentified cDNA clone (HGP-27) overexpressed in $PGE_2$-treated cells. The full-length cDNA of HGP-27 was isolated using RACE, which consisted of a 30-nt 5'-noncoding region, a 891-nt ORF encoding the 296 amino acid protein, and a 738-nt 3'-noncoding region including a poly(a) signal. This gene was localized on the short arm of chromosome number 11. Using the Motif Finder program, a myb-DNA binding repeat signature was detected on the ORF region. The COOH-terminal half was shown to have similarity with the $NH_3$-terminal domain of thioredoxin (Trx). This relation between HGP-27 and Trx implied a potential role for HGP-27 in modulating the DNA binding function of a transcription factor, myb.

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Nucleotide Sequence Analysis and Secondary Structure Modeling of the 3'-Noncoding Regions of Two Korean Strains of Turnip Mosaic Virus (순무 모자이크 바이러스 두 한국계통의 3' 말단 비번역부위에 대한 염기서열분석 및 2차구조 모델링)

  • 최장경;류기현;최국선;박원목
    • Korean Journal Plant Pathology
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    • v.11 no.3
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    • pp.271-277
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    • 1995
  • The RNA nucleotide sequences of the 3/-noncoding regions (3'-NCRs) of two Korean strains of turnip mosaic virus (TuMV), Ca and cqs, have been determined from their cDNA clones that encompassed the 3'-terminal regions of the viral genomic RNAs. The 3'-NCRs of both strains were 209 nucleotides long, terminated with GAC residues and poly (A) tails. The potential polyadenylational signal motif, UAUGU, was located 140 nucleotides upstream from the poly (A) tail in each of the virus. A highly conserved hexanucleotide sequence [A G U G A/U G/C], which was common in the 3'-NCRs of the potyvirus RNAs, was also found at the regions of 119 bases upstream from the 3'-end. Comparison of the 3'-NCRs of the two Korean isolates with those of four strains from Canada, China and Japan showed significantly identical genotypes (94.3∼99.5%). The secondary structure of three loops with long stems was found within the 3'-NCRs by sequence analysis. The substituted bases in the region among the six TuMV strains did not alter their secondary structures. Length of the 3'-NCRs of the know 11 potyviral RNAs and TuMV RNAs was different from one another and their nucleotide sequences showed 55.7% to 24.0% of homology. The 3'-NCR, therefore, is considered to be useful for phylogenetic studies in potyviruses.

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A Short Report on the Markov Property of DNA Sequences on 200-bp Genomic Units of ENCODE/Broad ChromHMM Annotations: A Computational Perspective

  • Park, Hyun-Seok
    • Genomics & Informatics
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    • v.16 no.3
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    • pp.65-70
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    • 2018
  • The non-coding DNA in eukaryotic genomes encodes a language which programs chromatin accessibility, transcription factor binding, and various other activities. The objective of this short report was to determine the impact of primary DNA sequence on the epigenomic landscape across 200-base pair genomic units by integrating nine publicly available ChromHMM Browser Extensible Data files of the Encyclopedia of DNA Elements (ENCODE) project. The nucleotide frequency profiles of nine chromatin annotations with the units of 200 bp were analyzed and integrative Markov chains were built to detect the Markov properties of the DNA sequences in some of the active chromatin states of different ChromHMM regions. Our aim was to identify the possible relationship between DNA sequences and the newly built chromatin states based on the integrated ChromHMM datasets of different cells and tissue types.