The helicase and RNaseIIIa domains of Arabidopsis Dicer-Like1 modulate catalytic parameters during MicroRNA biogenesis
KAUST DepartmentBiological and Environmental Sciences and Engineering (BESE) Division
Desert Agriculture Initiative
Online Publication Date2012-04-03
Print Publication Date2012-06-01
Permanent link to this recordhttp://hdl.handle.net/10754/562151
MetadataShow full item record
AbstractDicer-Like1 (DCL1), an RNaseIII endonuclease, and Hyponastic Leaves1 (HYL1), a double-stranded RNA-binding protein, are core components of the plant microRNA (miRNA) biogenesis machinery. hyl1 mutants accumulate low levels of miRNAs and display pleiotropic developmental phenotypes. We report the identification of five new hyl1 suppressor mutants, all of which are alleles of DCL1. These new alleles affect either the helicase or the RNaseIIIa domains of DCL1, highlighting the critical functions of these domains. Biochemical analysis of the DCL1 suppressor variants reveals that they process the primary transcript (pri-miRNA) more efficiently than wild-type DCL1, with both higher Kcat and lower Km values. The DCL1 variants largely rescue wild-type miRNA accumulation levels in vivo, but do not rescue the MIRNA processing precision defects of the hyl1 mutant. In vitro, the helicase domain confers ATP dependence on DCL1-catalyzed MIRNA processing, attenuates DCL1 cleavage activity, and is required for precise MIRNA processing of some substrates. © 2012 American Society of Plant Biologists.
CitationLiu, C., Axtell, M. J., & Fedoroff, N. V. (2012). The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis. Plant Physiology, 159(2), 748–758. doi:10.1104/pp.112.193508
SponsorsThis work was supported by the U.S. National Science Foundation (grant nos. 0718051 to M.J.A. and 0640186 to N.V.F.).
PubMed Central IDPMC3406889
- A dominant mutation in DCL1 suppresses the hyl1 mutant phenotype by promoting the processing of miRNA.
- Authors: Tagami Y, Motose H, Watanabe Y
- Issue date: 2009 Mar
- The interaction between DCL1 and HYL1 is important for efficient and precise processing of pri-miRNA in plant microRNA biogenesis.
- Authors: Kurihara Y, Takashi Y, Watanabe Y
- Issue date: 2006 Feb
- Complementation of HYPONASTIC LEAVES1 by double-strand RNA-binding domains of DICER-LIKE1 in nuclear dicing bodies.
- Authors: Liu Q, Yan Q, Liu Y, Hong F, Sun Z, Shi L, Huang Y, Fang Y
- Issue date: 2013 Sep
- The N-terminal double-stranded RNA binding domains of Arabidopsis HYPONASTIC LEAVES1 are sufficient for pre-microRNA processing.
- Authors: Wu F, Yu L, Cao W, Mao Y, Liu Z, He Y
- Issue date: 2007 Mar
- The RNA-binding proteins HYL1 and SE promote accurate in vitro processing of pri-miRNA by DCL1.
- Authors: Dong Z, Han MH, Fedoroff N
- Issue date: 2008 Jul 22