• 제목/요약/키워드: DNA mobility

검색결과 134건 처리시간 0.021초

The Regulation of AP-1 DNA Binding Activity by Long-term Nicotine Stimulation in Bovine Adrenal Medullary Chromaffin Cells: Role of Second Messengers

  • Lee, Jin-Koo;Choi, Seong-Soo;Suh, Hong-Won
    • The Korean Journal of Physiology and Pharmacology
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    • 제6권2호
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    • pp.109-112
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    • 2002
  • The signal pathways involved in the regulation of AP-1 DNA binding activity in long-term nicotine stimulated bovine adrenal medullary chromaffin (BAMC) cells have not been well characterized. To understand the involvement of second messengers in the regulation of AP-1 DNA binding activity, the present study was designed to define the time-course for inhibition of nicotine-induced responses by cholinergic antagonists, $Ca^{2+}$ and calmodulin (CaM) antagonists, and calcium/calmodulin-dependent protein kinase (CaMK) II inhibitor using electrophoretic mobility shift assay. Nicotine $(10{\mu}M)$ stimulation increased AP-1 DNA binding activity at 24 hr after treatment. Posttreatment with hexamethonium (1 mM) plus atropine $(1{\mu}M)$ (HA), nimodipine $(1{\mu}M),$ or calmidazolium $(1{\mu}M)$ at 0.5, 3, and 6 hr after the nicotine treatment significantly inhibited the AP-1 DNA binding activity increased by long-term nicotine stimulation. However, posttreatment with HA, nimodipine, or calmidazolium at 9 or 12 hr after the nicotine treatment did not affect the nicotine-induced increase of AP-1 DNA binding activity. The pretreatment of BAMC cells with various concentrations of KN-62 inhibited the increase of AP-1 DNA binding activity induced by nicotine in a concentration-dependent manner. KN-62 $(10{\mu}M)$ posttreatment beginning at 0.5, 3, or 6 hr after the nicotine treatment significantly inhibited the increase of AP-1 DNA binding activity. However, KN-62 posttreatment beginning at 9 or 12 hr after the nicotine treatment did not affect the increase of AP-1 DNA binding activity. This study suggested that stimulation (for at least 6 hr) of nicotinic receptors on BAMC cells was necessary for increase of AP-1 DNA binding activity, and activation of $Ca^{2+},$ CaM, and CaMK II up to 6 hr at least seemed to be required for the increase of nicotine-induced AP-1 DNA binding activity.

Suppression of AP-1 Activity by Tanshinone and Cancer Cell Growth Inhibition

  • 박세연;송지성;이덕근;양철학
    • Bulletin of the Korean Chemical Society
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    • 제20권8호
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    • pp.925-928
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    • 1999
  • The process of transcription is the major point at which gene expression is regulated. The jun and fos families of eukaryotic transcription factor heterodimerize to form complexes capable of binding 5'-TGAGTCA-3'DNA elements (AP-1 binding site). To search for the inhibitors of the jun-fos-DNA complex formation, several natural products extracts were screened and methanol extract of tanshen (the dried roots of Salvia miltiorrhiza Bunge) showed remarkable inhibitory activity. The active compounds of the extracts were purified using re-peated column chromatography and recrystallization. Their structures were identified as tanshinone I and tanshinone IIA. Through the electrophoresis mobility shift assay and cell cytotoxicity test, tanshinone I and tanshinone IIA were identified as inhibitors that suppress not only AP-1 function but also the cell proliferation. Tanshinone I also suppressed the jun-fos-DNA complex formation in TPA-induced NIH 3T3 cells.

Non-histone protein HMGB1 inhibits the repair of damaged DNA by cisplatin in NIH-3T3 murine fibroblasts

  • Yusein-Myashkova, Shazie;Ugrinova, Iva;Pasheva, Evdokia
    • BMB Reports
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    • 제49권2호
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    • pp.99-104
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    • 2016
  • The nuclear non-histone protein high mobility group box (HMGB) 1 is known to having an inhibitory effect on the repair of DNA damaged by the antitumor drug cisplatin in vitro. To investigate the role of HMGB1 in living cells, we studied the DNA repair of cisplatin damages in mouse fibroblast cell line, NIH-3T3. We evaluated the effect of the post-synthetic acetylation and C-terminal domain of the protein by overexpression of the parental and mutant GFP fused forms of HMGB1. The results revealed that HMGB1 had also an inhibitory effect on the repair of cisplatin damaged DNA in vivo. The silencing of HMGB1 in NIH-3T3 cells increased the cellular DNA repair potential. The increased levels of repair synthesis could be "rescued" and returned to less than normal levels if the knockdown cells were transfected with plasmids encoding HMGB1 and HMGB1 K2A. In this case, the truncated form of HMGB1 also exhibited a slight inhibitory effect.

Identification and Characterization of LHX8 DNA Binding Elements

  • Park, Miree;Jeon, Sanghyun;Jeong, Ji-Hye;Park, Miseon;Lee, Dong-Ryul;Yoon, Tae Ki;Choi, Dong Hee;Choi, Youngsok
    • 한국발생생물학회지:발생과생식
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    • 제16권4호
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    • pp.379-384
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    • 2012
  • Lhx8 (LIM homeobox 8) gene encodes a LIM homeodomain transcriptional regulator that is preferentially expressed in germ cells and critical for mammalian folliculogenesis. However, Lhx8 DNA binding sequences are not characterized yet. We aimed to identify and characterize a cis-acting sequence of germ-cell specific transcriptional factor, Lhx8. To identify Lhx8 DNA binding element, Cyclic Amplification of Sequence Target (CAST) Analysis was performed. Electrophoretic Mobility Shift Assay (EMSA) was processed for the binding specificity of Lhx8. Luciferase assay was for the transcriptional activity of Lhx8 through identified DNA binding site. We identified a putative cis-acting sequence, TGATTG as Lhx8 DNA binding element (LBE). In addition, Lhx8 binds to the LBE with high affinity and augments transcriptional activity of luciferase reporter driven by artificial promoter containing the Lhx8 binding element. These findings indicate that Lhx8 directly regulates the transcription of genes containing Lhx8 binding element in oocytes during early folliculogenesis.

Secondary structure analysis of MRA1997 from Mycobacterium tuberculosis and characterization of DNA binding property

  • Kim, Hyo Jung;Lee, Ki-Young;Kim, Yena;Kwon, Ae-Ran;Lee, Bong-Jin
    • 한국자기공명학회논문지
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    • 제20권2호
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    • pp.36-40
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    • 2016
  • MRA1997 is a highly conserved protein from mycobacterial strains. However, no structural and functional information is associated with it. Thus, to obtain details about structure and function of this protein, we have utilized NMR spectroscopy. The recombinant MRA1997 was highly purified and its DNA binding mode was characterized. The tertiary structure of MRA1997 was modeled on the basis of our NMR chemical shift data combined with the webserver CS23D. The binding of MRA1997 with DNA was first monitored by electrophoresis mobility shift assays. The residues involved in DNA binding are identified using NMR chemical shift perturbation experiments. Based on our study, we suggest that MRA1997 interacts with DNA and may play an important role in Mycobacterium tuberculosis physiology.

DNA (Data, Network, AI) 기반 지능형 정보 기술 (DNA (Data, Network, AI) Based Intelligent Information Technology)

  • 윤주상;한연희
    • 정보처리학회논문지:컴퓨터 및 통신 시스템
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    • 제9권11호
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    • pp.247-249
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    • 2020
  • 4차 산업혁명 시대에 다양한 분야에서 ICT 기술 간 융합에 대한 요구가 증가하고 있다. 이에 마쳐 데이터, 네트워크, 인공지능 기술이 결합한 새로운 용어인 DNA(Data, Network, AI)가 사용 중이다. DNA는 지능형 응용 및 서비스 개발에 있어 잠재적 기술력을 가지고 있다. 이에 본 논문에서는 DNA 기술 기반의 논리적 포그 네트워크 기반의 서비스 이미지 배치 기술, 산업용 무선 센서 네트워크에서의 기계학습 기반 이동성 기술, 뇌신호 주파수 특성을 이용한 CNN 기반 BCI 성능 예측 기술, 소스코드 주제를 이용한 인공신경망 기반 경고 분류 방법 기술, 챗봇 환경에서 데이터 시각화 인터랙션을 위한 자연어처리 기술에 대한 심사 완료된 논문들을 소개한다.

In vivo와 in vitro에서 DicA 단백질의 온도 의존적 DNA 결합 (Temperature-dependent DNA binding of DicA protein in vivo and in vitro)

  • 이연호;윤상훈;임헌만
    • 미생물학회지
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    • 제55권3호
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    • pp.181-190
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    • 2019
  • 대장균 세포분열 조절에 관여하는 DicA 단백질은 $37^{\circ}C$보다 $25^{\circ}C$에서 DNA에 더욱 잘 결합한다. 그러나 DicA 단백질의 온도의존적 DNA 결합에 대한 분자적 원인은 명확하지 않다. 본 연구에서는 DicA 단백질이 어떻게 DNA에 결합하며, 왜 온도 의존적 결합양상을 보이는지 알아보았다. In vivo DNA 결합 분석 결과 RovA나 SlyA와 같은 DicA의 상동성 단백질과는 달리 DicA는 N 말단에 있는 DNA 결합 도메인을 이용하여 20개의 염기쌍으로 이루어진 dicC 조절자 유전자(Oc)에 결합함을 보여주었다. 또한 in vivo 실험에서 DicA는 $37^{\circ}C$ 보다 $25^{\circ}C$에서 DNA에 더 잘 결합하는 것으로 알려진 Cnu 또는 H-NS의 영향을 받지 않고 자체적으로 Oc에서의 온도 의존적 DNA 결합을 보인다. 하지만 정제된 DicA 단백질을 이용한 in vitro binding 실험에서는 온도 의존적 DNA 결합이 관찰되지 않았다. Crude 단백질을 이용한 실험에서 DicA 단백질의 온도 의존적 DNA 결합이 관찰되는 것으로 보아 DicA의 온도 의존적 DNA (Oc) 결합은 crude 단백질내에 존재하는 아직 알려지지 않은 in vivo factor에 의해 일어난다.

Epidermal Growth Factor Receptor Gene Polymorphisms and Gastric Cancer in Iran

  • Abediankenari, Saeid;Jeivad, Fereshteh
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권5호
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    • pp.3187-3190
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    • 2013
  • Background: Epidermal growth factor receptor (EGFR) is a transmembrane receptor which contributes to many processes involved in cell survival, proliferation and inhibits apoptosis, that may lead to cancer development. Gastric cancer is one of the most common diseases of digestive system that has low 5-year-survival. The aim of this research was to determine the significance of EGFR tyrosine kinase domain gene polymorphisms in gastric cancer in Iran. Materials and Methods: In the present study, 83 patients with gastric cancer and 40 normal subjects were investigated for EGFR gene polymorphisms in exons 18-21 by PCR-SSCP. Then, DNA sequencing was conducted for different mobility shift bands. Finally the data were statistically analyzed using the chi-2 test and the SPSSver.16 program. Results: Exon 18 of EGFR gene showed three different bands in SSCP pattern and DNA sequencing displayed one mutation. SSCP pattern of Exons 19 and 21 did not show different migration bands. Exon 20 of EGFR gene revealed multiple migrate bands in SSCP pattern. DNA sequencing displayed 2 mutations in this exon: one mutation was caused amino acid change and another mutation was silent. Conclusion: It may be that EGFR tyrosine kinase gene polymorphisms differ between populations and screening could be useful in gastric cancer patients who might benefit from tyrosine kinase inhibitor therapy.

Intrinsic bent DNA colocalizes with the sequence involved in the Nd-sD mutation in the Bombyx mori fibroin light chain gene

  • Barbosa, Joice Felipes;Bravo, Juliana Pereira;Takeda, Karen Izumi;Zanatta, Daniela Bertolini;Silva, Jose Luis Da Conceicao;Balani, Valerio Americo;Fiorini, Adriana;Fernandez, Maria Aparecida
    • BMB Reports
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    • 제41권5호
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    • pp.394-399
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    • 2008
  • Multiple sequence alignments of the Bombyx mori fibroin light chain gene (fib-L) from hybrids and from Chinese and Japanese strains demonstrated that 51.6% of the fib-L third intron is conserved. One of these conserved segments, 41 bp long, contains the sequence CGTTATTATACATATT, which is duplicated in the B. mori Nd-$s^D$ mutant. In the present work, electrophoretic mobility assays and computational analyses revealed a major peak of intrinsic bent DNA within the segment that undergoes breakage in the previously-described Nd-$s^D$ mutation. This result suggested that this intrinsically-curved region might mediate DNA cleavage and enhance recombination events in the third intron of the Bombyx mori fib-L gene.