• Title/Summary/Keyword: RNA degradation intermediates

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MS2 Labeling of Endogenous Beta-Actin mRNA Does Not Result in Stabilization of Degradation Intermediates

  • Kim, Songhee H.;Vieira, Melissa;Kim, Hye-Jin;Kesawat, Mahipal Singh;Park, Hye Yoon
    • Molecules and Cells
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    • v.42 no.4
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    • pp.356-362
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    • 2019
  • The binding of MS2 bacteriophage coat protein (MCP) to MS2 binding site (MBS) RNA stem-loop sequences has been widely used to label mRNA for live-cell imaging at single-molecule resolution. However, concerns have been raised recently from studies with budding yeast showing aberrant mRNA metabolism following the MS2-GFP labeling. To investigate the degradation pattern of MS2-GFP-labeled mRNA in mammalian cells and tissues, we used Northern blot analysis of ${\beta}$-actin mRNA extracted from the Actb-MBS knock-in and $MBS{\times}MCP$ hybrid mouse models. In the immortalized mouse embryonic cell lines and various organ tissues derived from the mouse models, we found no noticeable accumulation of decay products of ${\beta}$-actin mRNA compared with the wild-type mice. Our results suggest that accumulation of MBS RNA decay fragments does not always happen depending on the mRNA species and the model organisms used.

Investigation of biodegradation pathway of dibenzofuran by Novosphingobium pentaromativorans US6-1 via transcriptomic and mass-spectrometric analysis (전사체와 대사물질 구조분석을 통한 Novosphingobium pentaromativorans US6-1의 dibenzofuran 분해 경로 해석)

  • Na, Hyeyun;Kwon, KaeKyoung
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.46-52
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    • 2018
  • Biodegradation pathway of dibenzofuran (DBF) of Novosphingobium pentaromativorans US6-1, a high-molecular-weight polycyclic aromatic hydrocarbons degrading strain, was investigated via analysis of metabolic intermediates and transcriptome. As a result, 3(2H)-benzofuranone, a basic skeleton of the metabolic intermediates produced by lateral dioxygenation process, was detected as an intermediate. RNA-Seq analysis confirmed that most of the expressed genes upon exposure to DBF were related to the lateral degradation pathway. Based on these results, the biodegradation pathway of DBF by N. pentaromativorans US6-1 was proposed.