• Title/Summary/Keyword: RNP

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Expression of PACT and EIF2C2, Implicated in RNAi and MicroRNA Pathways, in Various Human Cell Lines

  • Lee, Yong-Sun;Jeon, Yesu;Park, Jong-Hoon;Hwang, Deog-Su;Dutta, Anindya
    • Animal cells and systems
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    • v.8 no.3
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    • pp.213-220
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    • 2004
  • MicroRNA and siRNA (small interfering RNA), representative members of small RNA, exert their effects on target gene expression through association with protein complexes called miRNP (microRNA associated ribonucleoproteins) and RISC (RNA induced silencing complex), respectively. Although the protein complexes are yet to be fully characterized, human EIF2C2 protein has been identified as a component of both miRNP and RISC. In this report, we raised antiserum against EIF2C2 in order to begin understanding the protein complexes. An immunoblot result indicates that EIF2C2 protein is ubiquitously expressed in a variety of cell lines from human and mouse. EIF2C2 protein exists in both cellular compartments, as indicated by an immunoblot assay with a nuclear extract and a cytosolic fraction (S100 fraction) from HeLa S3 lysate. Depletion of EIF2C1 or EIF2C2 protein resulted in a decrease of microRNA, suggesting a possible role of these proteins in microRNA stability or biogenesis. We also prepared antiserum against dsRNA binding protein PACT, whose homologs in C. elegans and Drosophila are known to have a role in the RNAi (RNA interference) pathway. The expression of PACT protein was also observed in a wide range of cell lines.

Effects of the Repression of sphpr1 Expression on Growth and mRNA Export in Fission Yeast (분열효모에서 sphpr1 유전자의 결실이 생장 및 mRNA Export에 미치는 영향)

  • Lee, Hyun-Joo;Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.171-174
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    • 2012
  • THOC1/Hpr1 is one subunit of THO complex that is an evolutionally conserved assembly involved in the mRNP packaging and mRNA export during transcription elongation. In fission yeast Schizosaccharomyces pombe, an ortholog (spHpr1) of THOC1/Hpr1 was identified based on sequence alignment. A deletion mutant in a diploid strain was constructed by replacing one of spHpr1-coding region with a $kan^r$ gene using one-step gene disruption method. Tetrad analysis showed that the sphpr1 is essential for growth. Over-expression of sphpr1 from strong nmt1 promoter caused no defects of growth and mRNA export. However, repression of the sphpr1 expression resulted in growth inhibition accompanied by accumulation of poly$(A)^+$ RNA in the nucleus. These results suggest that spHpr1 is involved in mRNA export from the nucleus to cytoplasm.

A case of mixed connective tissue disease presenting initially with Raynaud's phenomenon (레이노드 증후군으로 초기 발현된 복합 교원성 질환 1예)

  • Kim, Soo Young;Choi, Young Seok;Kim, Young Ok;Woo, Young Jong
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.886-891
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    • 2008
  • Mixed connective tissue disease (MCTD) is characterized by diverse symptoms including rheumatoid arthritis, scleroderma, systemic lupus erythematosus, and dermatomyositis, associated with high titers of antibodies to extractable nuclear antigen (ENA), especially anti-ribonucleoprotein (anti-RNP) antibody. Since the first report of 25 cases with MCTD in adults, there have been only a few cases of MCTD reported in children. Here, we report a rare childhood case of MCTD in a 7-year-old girl presenting initially with Raynaud's phenomenon, swollen hands, and ulceration of the right index finger tip followed by alopecia and arthritis during follow-up.

Effects of spTho1 Deletion and Over-Expression on mRNA Export in Fission Yeast (분열효모에서 spTho1 유전자의 결실과 과발현이 생장 및 mRNA Export에 미치는 영향)

  • Cho, Ye-Seul;Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.401-404
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    • 2010
  • Tho1 is a RNA-binding protein that assembles co-transcriptionally onto the nascent mRNA and is thought to be involved in mRNP biogenesis and mature mRNA export to cytoplasm in budding yeast. In fission yeast Schizosaccharomyces pombe, a homologue of THO1 (spTho1) was identified based on sequence alignment. A deletion mutant in a diploid strain was constructed by replacing one of spTho1-coding region with an ura4+ gene using one-step gene disruption method. Tetrad analysis showed that the spTho1 was not essential for growth. The spTho1 mutant did not show any defects of bulk mRNA export. However, over-expression of spTho1 from strong nmt1 promoter caused the growth defects and accumulation of poly(A)$^+$ RNA in the nucleus. These results suggest that spTho1 is involved in mRNA export from the nucleus to cytoplasm though it is not essential.

Effects of spThoc7 Deletion on Growth and mRNA Export in Fission Yeast (분열효모에서 spThoc7 유전자의 결실이 생장 및 mRNA Export에 미치는 영향)

  • Koh, Eun-Jin;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.249-253
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    • 2014
  • THOC7/Mft1 is a member of the THO complex that is an evolutionally conserved assembly connecting transcription elongation with mRNP packaging and mRNA export. In fission yeast Schizosaccharomyces pombe, an ortholog (spThoc7) of THOC7/Mft1 was isolated by partial complementation of the lethality in synthetic lethal mutant, SLRsm1. A deletion mutant in a diploid strain was constructed by replacing one of spThoc7-coding region with an $ura4^+$ gene using one-step gene disruption method. Tetrad analysis showed that the spthoc7 is nonessential for growth. But the ${\Delta}thoc7$ null mutant showed slight defects of both growth and mRNA export. And the functional spThoc7-GFP protein is localized mainly in the nucleus. These results suggest that spThoc7 is also involved in mRNA export from the nucleus to cytoplasm.

Effects of Tho2, a component of THO complex, on growth and mRNA export in fission yeast (분열효모에서 THO 복합체의 구성요소인 Tho2가 생장 및 mRNA export에 미치는 영향)

  • Koh, Eun-Jin;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.181-185
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    • 2015
  • Tho2/THOC2 is a subunit of the THO complex that plays important roles in mRNP biogenesis connecting transcription with mRNA maturation and export. A fission yeast, Schizosaccharomyces pombe, ortholog of Tho2/THOC2 was identified from the genome database. Tetrad analysis showed that the S. pombe tho2 is essential for growth. Repression or overexpression of the tho2 gene caused growth defect with elongated cells, abnormal DNA distribution, and accumulation of $poly(A)^+$ RNA in the nucleus. And the functional GFP-Tho2 protein is localized mainly in the nucleus. Yeast two-hybrid analysis showed that Tho2 interacted with Tex1, another subunit of THO complex. These results suggest that S. pombe Tho2 is also involved in mRNA export from the nucleus and is a component of THO complex.

Partial Purification and Characterization of Enzymes Involved in the Processing of Pre-M1 RNA at the 3' End in Escherichia coli (대장균에서 선구-M1 RNA의 3'-말단 가공에 관여하는 효소들의 부분 정제와 그 특성 조사)

  • Kim, Ha Dong;Ko, Jae Hyeong;Cho, Bong Rae;Lee, Young Hoon;Park, In Won
    • Journal of the Korean Chemical Society
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    • v.43 no.3
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    • pp.307-314
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    • 1999
  • Ml RNA, the RNA component of RNase P from Escherichia coli, is produced by 3' processing of pre-Ml RNA, a major primary transcript of the rnpB gene. The enzyme fraction containing the processing activity was partially purified and characterized. Since exposure of the active fraction to the high salt condition results in the inactivation of the processing activity, the processing enzyme seems to be an enzyme complex composed of multiple enzymes. The enzyme fraction loses the processing activity when treated with the chemical nuclease lead(II) ion, but regains its activity by the addition of RNA isolated from the enzyme fraction itself, suggesting that an RNA molecule(s) may be essential for the processing activity. Analysis of cleavage sites produced by the partially purified enzyme fraction also implies that the 3' processing occurs by multiple enzymes and at least in two distinct pathways.

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Minimizing Energy Consumption of Sensor Networks with Energy Balance Ratio and Relay Node Placement (에너지 균형비와 중계노드 위치를 함께 고려한 센서 네트워크의 에너지 소비 최소화)

  • Sohn, Surg-Won;Han, Kwang-Rok
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.7
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    • pp.1549-1555
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    • 2009
  • The Relay node placement problem is one of the most important requirements for many wireless sensor networks because the lifetime of sensor networks is closely related with the placement of relay nodes which receive sensed data from sensor nodes and forward them to the base station. Relay node placement problem has focused at minimization of dissipated total energy of the sensor nodes in whole networks. However, minimum total energy causes the unbalance of consumed energy in sensor nodes due to different distances between relay nodes and sensor nodes. This paper proposes the concept of energy balance ratio and finds the locations of relay nodes using objective functions which maximize the energy balance ratio. Maximizing this ratio results in maximizing the network lifetime by minimizing the energy consumption of large-scale sensor networks. However, finding a solution to relay node placement problem is NP-hard and it is very difficult to get exact solutions. Therefore, we get approximate solutions to EBR-RNP problem which considers both energy balance ratio and relay node placement using constraint programming.

Evaluation of Combined Quantification of PCA3 and AMACR Gene Expression for Molecular Diagnosis of Prostate Cancer in Moroccan Patients by RT-qPCR

  • Maane, Imane Abdellaoui;El Hadi, Hicham;Qmichou, Zineb;Al Bouzidi, Abderrahmane;Bakri, Youssef;Sefrioui, Hassan;Dakka, Nadia;Moumen, Abdeladim
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.12
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    • pp.5229-5235
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    • 2016
  • Prostate cancer (PCa) remains one of the most widespread and perplexing of all human malignancies. Assessment of gene expression is thought to have an important impact on cancer diagnosis, prognosis and therapeutic decisions. In this context, we explored combined expression of PCa related target genes AMACR and PCA3 in 126 formalin fixed paraffin embedded prostate tissues (FFPE) from Moroccan patients, using quantitative real time reverse transcription-PCR (RT-qPCR). This quantification required data normalization accomplished using stably expressed reference genes (RGs). A panel of twelve RG was assessed, data being analyzed using GenEx V6 based on geNorm, NormFinder and statistical methods. Accordingly, the hnRNP A1 gene was identified and selected as the most stably expressed RG for reliable and accurate gene expression quantification in prostate tissues. The ratios of both PCA3 and AMACR gene expression relative to that of the hnRNP A1 gene were calculated and the performance of each target gene for PCa diagnosis was evaluated using receiver-operating characteristics. PCA3 and AMACR mRNA quantification based on RT-qPCR may prove useful in PCa diagnosis. Of particular interesting, combining PCA3 and AMACR quantification improved PCa prediction by increasing sensitivity with retention of good specificity.