|Table of Contents|

Identification of novel components of Arabidopsis miRNA pathway and mutant analysis(PDF)

《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

Issue:
2017年No.5(441-550)
Page:
464-470
Research Field:
生物工程
Publishing date:

Info

Title:
Identification of novel components of Arabidopsis miRNA pathway and mutant analysis
Author(s):
Zhao Qingzhe1 Liang Chao2 and Mo Beixin1
1) College of Life Sciences and Oceanography, Shenzhen University, Shenzhen Key Laboratory of Microbiology and Gene Engineering, Shenzhen 518060, Guangdong Province, P.R.China
2) College of Life Sciences and Oceanography, Shenzhen University, Guangdong Provincial Key Laboratory for Plant Epigenetics, Shenzhen 518060, Guangdong Province, P.R.China
Keywords:
molecular biology SUC2:amiR-SUL Arabidopsis thaliana forward genetic screening system microRNA (miRNA) AGO1 protein EMS mutagenesis quantitative real-time PCR(qRT-PCR) whole genome sequencing
PACS:
Q 946.2
DOI:
10.3724/SP.J.1249.2017.05464
Abstract:
Micro ribonucleic acids (miRNAs) are 20-24 nucleotides non-coding RNAs that play key regulatory roles in developmental and physiological processes in plants. In order to screen the new components that are involved in miRNA biogenesis, turnover and movement, we establish a forward genetic screening system using Arabidopsis thaliana transgenic line (SUC2:amiR-SUL). After ethylmethylsulfone (EMS) mutagenesis on SUC2:amiR-SUL, one stable mutant line (SUP-E45) is isolated. The result of phenotype observation, quantitative real-time polymerase chain reaction (qRT-PCR) and whole genome sequencing on the SUP-E45 mutant plants indicates that the altered phenotype is caused by the mutation in Argonaute 1 (Ago1) gene. AGO1 protein encoded by ago1 gene is crucial in the miRNA pathways, which recruits mature miRNA to form miRISC silence complex (miRNAs-induced silencing complex, miRISC) to negatively regulate the expression of target genes. The screening system can be used to select other factors that participate in the processing such as miRNA synthesis, miRNA’s activity or miRNA transport. The research can lay the foundation on the subsequent Arabidopsis miRNA pathway.

References:

[1] Chen Xuemei. Small RNAs and Their Roles in Plant Development[M]// Annual Review of Cell and Developmental Biology. 2009:21-44.
[2] 马轩,李盛本,莫蓓莘,等.拟南芥ago1-27突变体的RNA-seq分析[J].深圳大学学报理工版,2017,34(1):27-32.
Ma Xuan, Li Shengben, Mo Beixin, et al. RNA-seq analysis on Arabidopsis ago1-27 mutant[J]. Journal of Shenzhen University Science and Engineering, 2017, 34(1): 27-32.(in Chinese)
[3] Mallory A, Vaucheret H. Form, function, and regulation of ARGONAUTE proteins[J]. The Plant Cell, 2010, 22(12): 3879-3889.
[4] Ren Guodong, Xie Meng, Dou Yongchao, et al. Regulation of miRNA abundance by RNA binding protein TOUGH in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(31): 12817-12821.
[5] Han M H, Goud S, Song Liang, et al. The arabidopsis double-stranded RNA-binding protein HYL1 plays a role in microRNA-mediated gene regulation[J]. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(4): 1093-1098.
[6] Dong Zhicheng, Han M H, Fedoroff N. The RNA-binding proteins HYL1 and SE promote accurate in vitro processing of pri-miRNA by DCL1[J]. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105(29): 9970-9975.
[7] Park W, Li Junjie, Song Rentao, et al. CARPEL FACTORY, a dicer homolog, and HEN1, a novel protein, act in microRNA metabolism in arabidopsis thaliana[J]. Current Biology, 2002, 12(17): 1484-1495.
[8] Fang Yuda, Spector D L. Identification of nuclear dicing bodies containing proteins for microRNA biogenesis in living Arabidopsis plants[J]. Current Biology, 2007, 17(9): 818-823.
[9] Gandikota M, Birkenbihl R P, Hhmann S, et al. The miRNA156/157 recognition element in the 3′ UTR of the Arabidopsis SBP box gene SPL3 prevents early flowering by translational inhibition in seedlings [J]. The Plant Journal: for Cell and Molecular Biology, 2007, 49(4): 683-693.
[10] Chen Xuemei. A microRNA as a translational repressor of APETALA2 in Arabidopsis flower development[J]. Science, 2004, 303(5666): 2022-2025.
[11] Carlsbecker A, Lee J Y, Roberts C J, et al. Cell signalling by microRNA165/6 directs gene dose-dependent root cell fate[J]. Nature, 2010, 465(7296): 316-321.
[12] Shunsuke M, Koi S, Hashimoto T, et al. Non-cell-autonomous microRNA165 acts in a dose-dependent manner to regulate multiple differentiation status in the Arabidopsis root[J]. Development, 2011, 138(11): 2303-2313.
[13] Chiou T J, Aung K, Lin Shui, et al. Regulation of phosphate homeostasis by MicroRNA in Arabidopsis[J]. The Plant Cell, 2006, 18(2): 412-421.
[14] Buhtz A, Springer F, Chappell L, et al. Identification and characterization of small RNAs from the phloem of Brassica napus[J]. The Plant Journal: for Cell and Molecular Biology, 2008, 53(5): 739-749.
[15] Martin A, Adam H, Díaz-Mendoza M, et al. Graft-transmissible induction of potato tuberization by the microRNA miR172[J]. Development, 2009, 136(17): 2873-2881.
[16] 方晓峰. 拟南芥MicroRNA通路新因子的鉴定和作用机制研究[D]. 北京:北京协和医学院, 2014.
Fang Xiaofeng. Identification and Characterization of Novel Components in the Arabidopsis MicroRNA Pathway[D]. Beijing: Peking Union Medical College,2014.(in Chinese)
[17] Morel J B, Godon C, Mourrain P, et al. Fertile hypomorphic ARGONAUTE (ago1) mutants impaired in post-transcriptional gene silencing and virus resistance[J]. The Plant Cell, 2002, 14(3): 629-639.
[18] Zhang Shuxin, Xie Meng, Ren Guodong, et al. CDC5, a DNA binding protein, positively regulates posttranscriptional processing and/or transcription of primary microRNA transcript[J]. Proceedings of the National Academy of Sciences of the United States of America, 2013, 110(43): 17588-17593.
[19] Wu Xueying, Shi Yupeng, Li Jingrui, et al. A role for the RNA-binding protein MOS2 in microRNA maturation in Arabidopsis[J]. Cell Research, 2013, 23(5): 645-657.
[20] 岳路明,宋剑波,莫蓓莘,等.拟南芥AGO基因家族分析及盐胁迫下的表达验证[J].深圳大学学报理工版,2017,34(4):331-440.
Yue Luming, Song Jianbo, Mo Beixin,et al. Bioinformatical and experimental analysis of AGO genes in response to salt stress[J]. Journal of Shenzhen University Science and Engineering,2017, 34(4): 331-440.(in Chinese)

Memo

Memo:
-
Last Update: 2017-09-11