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MicroR159 regulation of most conserved targets in Arabidopsis has negligible phenotypic effects

Allen, Robert S; Li, Junyan; Alonso-Peral, Maria M; White, Rosemary G; Gubler, Frank; Millar, Anthony A

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BACKGROUND A current challenge of microRNA (miRNA) research is the identification of biologically relevant miRNA:target gene relationships. In plants, high miRNA:target gene complementarity has enabled accurate target predictions, and slicing of target mRNAs has facilitated target validation through rapid amplification of 5' cDNA ends (5'-RACE) analysis. Together, these approaches have identified more than 20 targets potentially regulated by the deeply conserved miR159 family in Arabidopsis,...[Show more]

dc.contributor.authorAllen, Robert S
dc.contributor.authorLi, Junyan
dc.contributor.authorAlonso-Peral, Maria M
dc.contributor.authorWhite, Rosemary G
dc.contributor.authorGubler, Frank
dc.contributor.authorMillar, Anthony A
dc.date.accessioned2016-01-21T00:50:06Z
dc.date.available2016-01-21T00:50:06Z
dc.identifier.issn1758-907X
dc.identifier.urihttp://hdl.handle.net/1885/95604
dc.description.abstractBACKGROUND A current challenge of microRNA (miRNA) research is the identification of biologically relevant miRNA:target gene relationships. In plants, high miRNA:target gene complementarity has enabled accurate target predictions, and slicing of target mRNAs has facilitated target validation through rapid amplification of 5' cDNA ends (5'-RACE) analysis. Together, these approaches have identified more than 20 targets potentially regulated by the deeply conserved miR159 family in Arabidopsis, including eight MYB genes with highly conserved miR159 target sites. However, genetic analysis has revealed the functional specificity of the major family members, miR159a and miR159b is limited to only two targets, MYB33 and MYB65. Here, we examine the functional role of miR159 regulation for the other potential MYB target genes. RESULTS For these target genes, functional analysis failed to identify miR159 regulation that resulted in any major phenotypic impact, either at the morphological or molecular level. This appears to be mainly due to the quiescent nature of the remaining family member, MIR159c. Although its expression overlaps in a temporal and spatial cell-specific manner with a subset of these targets in anthers, the abundance of miR159c is extremely low and concomitantly a mir159c mutant displays no anther defects. Examination of potential miR159c targets with conserved miR159 binding sites found neither their spatial or temporal expression domains appeared miR159 regulated, despite the detection of miR159-guided cleavage products by 5'-RACE. Moreover, expression of a miR159-resistant target (mMYB101) resulted predominantly in plants that are indistinguishable from wild type. Plants that displayed altered morphological phenotypes were found to be ectopically expressing the mMYB101 transgene, and hence were misrepresentative of the in vivo functional role of miR159. CONCLUSIONS This study presents a novel explanation for a paradox common to plant and animal miRNA systems, where among many potential miRNA-target relationships usually only a few appear physiologically relevant. The identification of a quiescent miR159c:target gene regulatory module in anthers provides a likely rationale for the presence of conserved miR159 binding sites in many targets for which miR159 regulation has no obvious functional role. Remnants from the demise of such modules may lead to an overestimation of miRNA regulatory complexity when investigated using bioinformatic, 5'-RACE or transgenic approaches.
dc.description.sponsorshipRSA was funded by an ANU postgraduate scholarship and by a CSIRO Emerging Science Initiative. JL is the recipient of an ANU international student postgraduate scholarship. This research was supported by an Australian Research Council grant DP0773270.
dc.publisherBioMed Central
dc.rights© 2010 Allen et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
dc.sourceSilence
dc.subjectKeywords: microRNA; Arabidopsis; article; binding site; cell specificity; controlled study; gene control; gene expression; gene targeting; in vivo study; mutant; nonhuman; phenotype; priority journal; transgene; wild type; Animalia; Arabidopsis
dc.titleMicroR159 regulation of most conserved targets in Arabidopsis has negligible phenotypic effects
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume1
dc.date.issued2010-10-28
local.identifier.absfor060705
local.identifier.absfor060702
local.identifier.ariespublicationu4956746xPUB48
local.publisher.urlhttp://www.biomedcentral.com/
local.type.statusPublished Version
local.contributor.affiliationAllen, Robert, College of Medicine, Biology and Environment, CMBE Research School of Biology, Division of Plant Sciences, The Australian National University
local.contributor.affiliationLi, Jun, College of Medicine, Biology and Environment, CMBE Research School of Biology, Division of Biomedical Science and Biochemistry, The Australian National University
local.contributor.affiliationAlonso Peral, Maria, College of Medicine, Biology and Environment, CMBE Research School of Biology, Division of Plant Sciences, The Australian National University
local.contributor.affiliationWhite, Rosemary Gillian, CSIRO Division of Plant Industry, Australia
local.contributor.affiliationGubler, Frank , CSIRO, Australia
local.contributor.affiliationMillar, Anthony, College of Medicine, Biology and Environment, CMBE Research School of Biology, Division of Plant Sciences, The Australian National University
dc.relationhttp://purl.org/au-research/grants/arc/DP0773270
local.identifier.essn1758-907X
local.bibliographicCitation.issue1
local.bibliographicCitation.startpage18
local.identifier.doi10.1186/1758-907X-1-18
local.identifier.absseo829999
local.identifier.absseo970106
dc.date.updated2016-02-24T11:28:56Z
local.identifier.scopusID2-s2.0-79952334906
CollectionsANU Research Publications

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