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Plant and Cell Physiology Advance Access originally published online on August 3, 2007
Plant and Cell Physiology 2007 48(9):1243-1253; doi:10.1093/pcp/pcm099
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Rapid Paper

Location of a Possible miRNA Processing Site in SmD3/SmB Nuclear Bodies in Arabidopsis

Yoichiro Fujioka1, Maki Utsumi1, Yusuke Ohba2 and Yuichiro Watanabe1,*

1Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Komaba 3-8-1, Meguro, Tokyo, 153-8902 Japan
2Laboratory of Pathophysiology and Signal Transduction, Department of Pathology, Hokkaido University Graduate School of Medicine, N15 W7, Kita-ku, Sapporo, 060-8638 Japan

*Corresponding author: E-mail, solan{at}bio.c.u-tokyo.ac.jp; Fax, +81-3-5454-6776.


  Abstract

There has been much recent research on the contribution of microRNA (miRNA) in plant organogenesis and hormone action. In plants, it has been reported that Dicer-like 1 (DCL1), HYPONASTIC LEAVES1 (HYL1) and SERRATE (SE) are involved in the production of miRNAs. The means by which miRNAs are processed and transported is not well understood in detail, however. In this study, we investigated the intracellular localization and intermolecular interaction of these molecules using imaging techniques, including bimolecular fluorescence complementation and fluorescence resonance energy transfer techniques, making use of various enhanced fluorescent proteins. We found that DCL1, HYL1 and SE formed bodies which localized in the nuclei. We were also able to locate the miRNA primary transcript using an MS2-tagged method on these bodies. It appears very likely that the observed DCL1–HYL1–SE nuclear body is involved in miRNA production. Co-expression of SmD3 or SmB proteins revealed the localization of DCL1–HYL1–SE complexes in the SmD3/SmB nuclear bodies.

Keywords: Arabidopsis - Dicer-like 1 - HYL1 - Micro RNA - Processing - SERRATE

Abbreviations: BiFC, bimolecular fluorescence complementation; CB, Cajal body; DCL1, DICER-LIKE 1; DEX, dexamethasone; dsRNA, double-stranded RNA; FRET, fluorescence resonance energy transfer; HA, hemagglutinin; HYL1, HYPONASTIC LEAVES1; miRNA, micro RNA; mRFP, monomeric red fluorescent protein; pre-miRNA, miRNA precursor; pri-miRNA, miRNA primary transcript; SE, SERRATE; SECFP, super-enhanced cyan fluorescent protein; siRNA, small interfering RNA; SmD3, small nuclear RNA-binding protein D3; YFP, yellow fluorescent protein.

(Received July 11, 2007; Accepted July 25, 2007)
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