Pre-microRNA processing activity in nuclear extracts from Arabidopsis suspension cells

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• 作者：Manabu Yoshikawa
• 关键词：Arabidopsis thaliana ; DICER ; LIKE1 ; MicroRNA ; Nuclear extract ; Pre ; miRNA
• 刊名：Journal of Plant Research
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：130
• 期：1
• 页码：75-82
• 全文大小：
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Plant Sciences; Plant Ecology; Plant Physiology; Plant Biochemistry;
• 出版者：Springer Japan
• ISSN：1618-0860
• 卷排序：130

文摘

MicroRNAs (miRNAs) play important roles in a variety of biological phenomena, such as development and responses to abiotic and biotic stresses, by regulating complementary target transcripts. Primary MIRNA transcripts (pri-miRNAs) are processed into miRNA/miRNA* duplexes via pre-miRNA intermediates by DICER family proteins. In plants miRNA/miRNA* duplexes are produced from pri-miRNAs exclusively by a complex containing DICER-LIKE1 (DCL1), Hyponastic leaves1 (HYL1) and SERREATE (SE) in the nucleus. Pri-miRNA or pre-miRNA processing activity has been detected using recombinant DCL1, HYL1 and SE proteins expressed in insect cells and immunopurified DCL1-HYL1-SE-containing complexes that transiently co-overexpressed in Nicotiana benthamiana. However, the processing of pre-miRNAs into miRNA/miRNA* using nuclear extracts has not been reported. Here I report the detection of pre-miRNA processing in nuclear extracts prepared from Arabidopsis suspension cells. In the nuclear extracts including DCL1, small RNAs that were 21 nucleotides in length were excised from a part of miRNA/miRNA* regions on pre-miRNAs. This system is potentially useful for in vitro analyses of pre-miRNA processing.