The helicase and RNaseIIIa domains of Arabidopsis Dicer-Like1 modulate catalytic parameters during MicroRNA biogenesis

Handle URI:
http://hdl.handle.net/10754/562151
Title:
The helicase and RNaseIIIa domains of Arabidopsis Dicer-Like1 modulate catalytic parameters during MicroRNA biogenesis
Authors:
Liu, Chenggang; Axtell, Michael J.; Fedoroff, Nina V.
Abstract:
Dicer-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.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division
Publisher:
American Society of Plant Biologists
Journal:
Plant Physiology
Issue Date:
3-Apr-2012
DOI:
10.1104/pp.112.193508
PubMed ID:
22474216
PubMed Central ID:
PMC3406889
Type:
Article
ISSN:
00320889
Sponsors:
This work was supported by the U.S. National Science Foundation (grant nos. 0718051 to M.J.A. and 0640186 to N.V.F.).
Additional Links:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3406889
Appears in Collections:
Articles; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorLiu, Chenggangen
dc.contributor.authorAxtell, Michael J.en
dc.contributor.authorFedoroff, Nina V.en
dc.date.accessioned2015-08-03T09:45:59Zen
dc.date.available2015-08-03T09:45:59Zen
dc.date.issued2012-04-03en
dc.identifier.issn00320889en
dc.identifier.pmid22474216en
dc.identifier.doi10.1104/pp.112.193508en
dc.identifier.urihttp://hdl.handle.net/10754/562151en
dc.description.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.en
dc.description.sponsorshipThis work was supported by the U.S. National Science Foundation (grant nos. 0718051 to M.J.A. and 0640186 to N.V.F.).en
dc.publisherAmerican Society of Plant Biologistsen
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3406889en
dc.titleThe helicase and RNaseIIIa domains of Arabidopsis Dicer-Like1 modulate catalytic parameters during MicroRNA biogenesisen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalPlant Physiologyen
dc.identifier.pmcidPMC3406889en
dc.contributor.institutionDepartment of Biology, Pennsylvania State University, University Park, PA 16802, United Statesen
dc.contributor.institutionPlant Biology Division, Samuel Roberts Noble Foundation, Ardmore, OK 73401, United Statesen
kaust.authorFedoroff, Nina V.en

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