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1.
Proc Natl Acad Sci U S A ; 111(26): 9407-12, 2014 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-24979769

RESUMEN

Auxin influences nearly every aspect of plant biology through a simple signaling pathway; however, it remains unclear how much of the diversity in auxin effects is explained by variation in the core signaling components and which properties of these components may contribute to diversification in response dynamics. Here, we recapitulated the entire Arabidopsis thaliana forward nuclear auxin signal transduction pathway in Saccharomyces cerevisiae to test whether signaling module composition enables tuning of the dynamic response. Sensitivity analysis guided by a small mathematical model revealed the centrality of auxin/indole-3-acetic acid (Aux/IAA) transcriptional corepressors in controlling response dynamics and highlighted the strong influence of natural variation in Aux/IAA degradation rates on circuit performance. When the basic auxin response circuit was expanded to include multiple Aux/IAAs, we found that dominance relationships between coexpressed Aux/IAAs were sufficient to generate distinct response modules similar to those seen during plant development. Our work provides a new method for dissecting auxin signaling and demonstrates the key role of Aux/IAAs in tuning auxin response dynamics.


Asunto(s)
Arabidopsis/fisiología , Ácidos Indolacéticos/metabolismo , Modelos Biológicos , Transducción de Señal/fisiología , Arabidopsis/metabolismo , Citometría de Flujo , Vectores Genéticos/genética , Microscopía Fluorescente , Saccharomyces cerevisiae , Biología Sintética
2.
Plant Physiol ; 160(1): 135-42, 2012 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-22843664

RESUMEN

Explaining how the small molecule auxin triggers diverse yet specific responses is a long-standing challenge in plant biology. An essential step in auxin response is the degradation of Auxin/Indole-3-Acetic Acid (Aux/IAA, referred to hereafter as IAA) repressor proteins through interaction with auxin receptors. To systematically characterize diversity in degradation behaviors among IAA|receptor pairs, we engineered auxin-induced degradation of plant IAA proteins in yeast (Saccharomyces cerevisiae). We found that IAA degradation dynamics vary widely, depending on which receptor is present, and are not encoded solely by the degron-containing domain II. To facilitate this and future studies, we identified a mathematical model able to quantitatively describe IAA degradation behavior in a single parameter. Together, our results demonstrate the remarkable tunability conferred by specific configurations of the auxin response pathway.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas F-Box/metabolismo , Ácidos Indolacéticos/metabolismo , Receptores de Superficie Celular/metabolismo , Transducción de Señal , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas F-Box/genética , Citometría de Flujo , Semivida , Ácidos Indolacéticos/farmacología , Modelos Biológicos , Reguladores del Crecimiento de las Plantas/metabolismo , Plantas Modificadas Genéticamente/efectos de los fármacos , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Estructura Terciaria de Proteína , Proteolisis , Receptores de Superficie Celular/genética , Proteínas Represoras/metabolismo , Saccharomyces cerevisiae/efectos de los fármacos , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Especificidad de la Especie , Factores de Tiempo , Transformación Genética , Ubiquitinación
3.
Genetics ; 186(4): 1247-60, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-20923975

RESUMEN

Slk19p is a member of the Cdc-14 early anaphase release (FEAR) pathway, a signaling network that is responsible for activation of the cell-cycle regulator Cdc14p in Saccharomyces cerevisiae. Disruption of the FEAR pathway results in defects in anaphase, including alterations in the assembly and behavior of the anaphase spindle. Many phenotypes of slk19Δ mutants are consistent with a loss of FEAR signaling, but other phenotypes suggest that Slk19p may have FEAR-independent roles in modulating the behavior of microtubules in anaphase. Here, a series of SLK19 in-frame deletion mutations were used to test whether Slk19p has distinct roles in anaphase that can be ascribed to specific regions of the protein. Separation-of-function alleles were identified that are defective for either FEAR signaling or aspects of anaphase spindle function. The data suggest that in early anaphase one region of Slk19p is essential for FEAR signaling, while later in anaphase another region is critical for maintaining the coordination between spindle elongation and the growth of interpolar microtubules.


Asunto(s)
Anafase , Proteínas Asociadas a Microtúbulos/fisiología , Proteínas de Saccharomyces cerevisiae/fisiología , Huso Acromático , Alelos , Proteínas de Ciclo Celular/metabolismo , Microtúbulos , Proteínas Tirosina Fosfatasas/metabolismo , Saccharomyces cerevisiae/citología , Proteínas de Saccharomyces cerevisiae/metabolismo , Transducción de Señal
4.
Virology ; 369(2): 234-44, 2007 Dec 20.
Artículo en Inglés | MEDLINE | ID: mdl-17825342

RESUMEN

The genome sequence of the Salmonella enterica serovar Anatum-specific, serotype-converting bacteriophage epsilon15 has been completed. The nonredundant genome contains 39,671 bp and 51 putative genes. It most closely resembles the genome of phiV10, an Escherichia coli O157:H7-specific temperate phage, with which it shares 36 related genes. More distant relatives include the Burkholderia cepacia-specific phage, BcepC6B (8 similar genes), the Bordetella bronchiseptica-specific phage, BPP-1 (8 similar genes) and the Photobacterium profundum prophage, P Pphipr1 (6 similar genes). epsilon15 gene identifications based on homologies with known gene families include the terminase small and large subunits, integrase, endolysin, two holins, two DNA methylase enzymes (one adenine-specific and one cytosine-specific) and a RecT-like enzyme. Genes identified experimentally include those coding for the serotype conversion proteins, the tail fiber, the major capsid protein and the major repressor. epsilon15's attP site and the Salmonella attB site with which it interacts during lysogenization have also been determined.


Asunto(s)
Genoma Viral , Fagos de Salmonella/genética , Salmonella enterica/virología , Secuencia de Aminoácidos , Tipificación de Bacteriófagos , Secuencia de Bases , ADN Bacteriano/genética , ADN Viral/genética , Datos de Secuencia Molecular , Fagos de Salmonella/clasificación , Fagos de Salmonella/fisiología , Salmonella enterica/clasificación , Salmonella enterica/genética , Homología de Secuencia de Ácido Nucleico , Serotipificación , Especificidad de la Especie , Proteínas Virales/genética , Ensamble de Virus , Integración Viral
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