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1.
New Phytol ; 188(1): 23-9, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20649916

ABSTRACT

• BREVIS RADIX (BRX) has been identified through a loss-of-function allele in the Umkirch-1 accession in a natural variation screen for Arabidopsis root growth vigor. Physiological and gene expression analyses have suggested that BRX is rate limiting for auxin-responsive gene expression by mediating cross-talk with the brassinosteroid pathway, as impaired root growth and reduced auxin perception of brx can be (partially) rescued by external brassinosteroid application. • Using genetic tools, we show that brx mutants also display significantly reduced cotyledon and leaf growth. • Similar to the root, the amplitude and penetrance of this phenotype depends on genetic background and shares the physiological features, reduced auxin perception and brassinosteroid rescue. Furthermore, reciprocal grafting experiments between mutant and complemented brx shoot scions and root stocks suggest that the shoot phenotypes are not an indirect consequence of the root phenotype. Finally, BRX gain-of-function lines display epinastic leaf growth and, in the case of dominant negative interference, increased epidermal cell size. Consistent with an impact of BRX on brassinosteroid biosynthesis, this phenotype is accompanied by increased brassinosteroid levels. • In summary, our results demonstrate a ubiquitous, although quantitatively variable role of BRX in modulating the growth rate in both the root and shoot.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/growth & development , Plant Shoots/growth & development , Arabidopsis/cytology , Arabidopsis/metabolism , Brassinosteroids , Cholestanols/metabolism , Cotyledon/anatomy & histology , Cotyledon/cytology , Cotyledon/metabolism , Mutation/genetics , Organ Size , Phenotype , Plant Leaves/anatomy & histology , Plant Leaves/growth & development , Plant Leaves/metabolism , Plant Roots/growth & development , Plant Roots/metabolism , Plant Shoots/cytology , Plant Shoots/metabolism , Steroids, Heterocyclic/metabolism
2.
Proc Natl Acad Sci U S A ; 107(18): 8475-80, 2010 May 04.
Article in English | MEDLINE | ID: mdl-20404146

ABSTRACT

Quantitative trait loci analysis of natural Arabidopsis thaliana accessions is increasingly exploited for gene isolation. However, to date this has mostly revealed deleterious mutations. Among them, a loss-of-function allele identified the root growth regulator BREVIS RADIX (BRX). Here we present evidence that BRX and the paralogous BRX-LIKE (BRXL) genes are under selective constraint in monocotyledons as well as dicotyledons. Unexpectedly, however, whereas none of the Arabidopsis orthologs except AtBRXL1 could complement brx null mutants when expressed constitutively, nearly all monocotyledon BRXLs tested could. Thus, BRXL proteins seem to be more diversified in dicotyledons than in monocotyledons. This functional diversification was correlated with accelerated rates of sequence divergence in the N-terminal regions. Population genetic analyses of 30 haplotypes are suggestive of an adaptive role of AtBRX and AtBRXL1. In two accessions, Lc-0 and Lov-5, seven amino acids are deleted in the variable region between the highly conserved C-terminal, so-called BRX domains. Genotyping of 42 additional accessions also found this deletion in Kz-1, Pu2-7, and Ws-0. In segregating recombinant inbred lines, the Lc-0 allele (AtBRX(Lc-0)) conferred significantly enhanced root growth. Moreover, when constitutively expressed in the same regulatory context, AtBRX(Lc-0) complemented brx mutants more efficiently than an allele without deletion. The same was observed for AtBRXL1, which compared with AtBRX carries a 13 amino acid deletion that encompasses the deletion found in AtBRX(Lc-0). Thus, the AtBRX(Lc-0) allele seems to contribute to natural variation in root growth vigor and provides a rare example of an experimentally confirmed, hyperactive allelic variant.


Subject(s)
Alternative Splicing , Arabidopsis Proteins/genetics , Arabidopsis/genetics , Plant Roots/genetics , Quantitative Trait Loci , Alleles , Amino Acid Sequence , Arabidopsis/chemistry , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/metabolism , Evolution, Molecular , Gene Deletion , Haplotypes , Molecular Sequence Data , Phylogeny , Plant Roots/growth & development , Plant Roots/metabolism , Sequence Alignment
3.
Development ; 136(12): 2059-67, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19465596

ABSTRACT

In Arabidopsis, interplay between nuclear auxin perception and trans-cellular polar auxin transport determines the transcriptional auxin response. In brevis radix (brx) mutants, this response is impaired, probably indirectly because of disturbed crosstalk between the auxin and brassinosteroid pathways. Here we provide evidence that BRX protein is plasma membrane-associated, but translocates to the nucleus upon auxin treatment to modulate cellular growth, possibly in conjunction with NGATHA class B3 domain-type transcription factors. Application of the polar auxin transport inhibitor naphthalene phthalamic acid (NPA) resulted in increased BRX abundance at the plasma membrane. Thus, nuclear translocation of BRX could depend on cellular auxin concentration or on auxin flux. Supporting this idea, NPA treatment of wild-type roots phenocopied the brx root meristem phenotype. Moreover, BRX is constitutively turned over by the proteasome pathway in the nucleus. However, a stabilized C-terminal BRX fragment significantly rescued the brx root growth phenotype and triggered a hypocotyl gain-of-function phenotype, similar to strong overexpressors of full length BRX. Therefore, although BRX activity is required in the nucleus, excess activity interferes with normal development. Finally, similar to the PIN-FORMED 1 (PIN1) auxin efflux carrier, BRX is polarly localized in vascular cells and subject to endocytic recycling. Expression of BRX under control of the PIN1 promoter fully rescued the brx short root phenotype, suggesting that the two genes act in the same tissues. Collectively, our results suggest that BRX might provide a contextual readout to synchronize cellular growth with the auxin concentration gradient across the root tip.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Cell Membrane/metabolism , Cell Nucleus/metabolism , Indoleacetic Acids/pharmacology , Arabidopsis/embryology , Arabidopsis Proteins/genetics , Endocytosis , Gene Expression Regulation, Plant , Indoleacetic Acids/metabolism , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Mutation , Plant Roots/embryology , Plant Roots/metabolism , Protein Transport
4.
J Nat Prod ; 68(12): 1793-5, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16378379

ABSTRACT

A new quaternary beta-carboline alkaloid, nostocarboline, was isolated from the freshwater cyanobacterium Nostoc 78-12A, and its constitution was assigned by 2D-NMR methods. The structure was proven by its total synthesis starting from norharmane via chlorination at C-6 and methylation at N-2. Nostocarboline was found to be a potent butyrylcholinesterase (BChE) inhibitor, with an IC(50) of 13.2 microM. The related 2-methylnorharmane, which is present in the human brain and might be relevant to Parkinson's disease (PD), was also determined to be a BChE inhibitor (11.2 microM). These inhibitory concentrations are comparable to galanthamine, an approved drug for the treatment of Alzheimer's disease (AD). Nostocarboline can thus be considered as a lead for the development of novel neurochemicals.


Subject(s)
Butyrylcholinesterase/metabolism , Carbolines , Cholinesterase Inhibitors , Galantamine/pharmacology , Nostoc/chemistry , Brain/enzymology , Cholinesterase Inhibitors/chemistry , Cholinesterase Inhibitors/isolation & purification , Cholinesterase Inhibitors/pharmacology , Humans , Inhibitory Concentration 50 , Molecular Structure , Parkinson Disease/drug therapy
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