RESUMEN
The plant-specific TOPLESS (TPL) family of transcriptional corepressors is integral to multiple angiosperm developmental processes. Despite this, we know little about TPL function in other plants. To address this gap, we investigated the roles TPL plays in the bryophyte Physcomitrium patens, which diverged from angiosperms approximately 0.5 billion years ago. Although complete loss of PpTPL function is lethal, transgenic lines with reduced PpTPL activity revealed that PpTPLs are essential for two fundamental developmental switches in this plant: the transitions from basal photosynthetic filaments (chloronemata) to specialised foraging filaments (caulonemata) and from two-dimensional (2D) to three-dimensional (3D) growth. Using a transcriptomics approach, we integrated PpTPL into the regulatory network governing 3D growth and we propose that PpTPLs represent another important class of regulators that are essential for the 2D-to-3D developmental switch. Transcriptomics also revealed a previously unknown role for PpTPL in the regulation of flavonoids. Intriguingly, 3D growth and the formation of caulonemata were crucial innovations that facilitated the colonisation of land by plants, a major transformative event in the history of life on Earth. We conclude that TPL, which existed before the land plants, was co-opted into new developmental pathways, enabling phytoterrestrialisation and the evolution of land plants.
Asunto(s)
Bryopsida , Plantas , Proteínas Co-Represoras/metabolismo , Plantas/metabolismo , Factores de Transcripción/metabolismo , Bryopsida/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
The regulation of protein synthesis plays an important role in the growth and development of all organisms. Upstream open reading frames (uORFs) are commonly found in eukaryotic messenger RNA transcripts and typically attenuate the translation of associated downstream main ORFs (mORFs). Conserved peptide uORFs (CPuORFs) are a rare subset of uORFs, some of which have been shown to conditionally regulate translation by ribosome stalling. Here, we show that Arabidopsis CPuORF19, CPuORF46 and CPuORF47, which are ancient in origin, regulate translation of any downstream ORF, in response to the agriculturally significant environmental signals, heat stress and water limitation. Consequently, these CPuORFs represent a versatile toolkit for inducible gene expression with broad applications. Finally, we note that different classes of CPuORFs may operate during distinct phases of translation, which has implications for the bioengineering of these regulatory factors.
Asunto(s)
Arabidopsis , Ribosomas , Arabidopsis/genética , Sistemas de Lectura Abierta/genética , Péptidos/genética , Plantas/genética , Biosíntesis de Proteínas , Ribosomas/genéticaRESUMEN
Transcriptional corepressors play important roles in establishing the appropriate levels of gene expression during growth and development. The TOPLESS (TPL) family of corepressors are critical for all plant life. TPLs are involved in numerous developmental processes and in the response to extrinsic challenges. As such these proteins have been the focus of intense study since Long and colleagues first described the TPL corepressor in 2006. In this review we will explore the evolutionary history of these essential plant-specific proteins, their mechanism of action based on recent structural analyses, and the myriad of pathways in which they function. We speculate how relatively minor changes in the peptide sequence of transcriptional regulators allowed them to recruit TPL into new processes, driving innovation and resulting in TPL becoming vital for plant development.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas Co-Represoras/metabolismo , Expresión Génica , Regulación de la Expresión Génica de las PlantasRESUMEN
Nonsense-mediated mRNA decay (NMD) is important for RNA quality control and gene regulation in eukaryotes. NMD targets aberrant transcripts for decay and also directly influences the abundance of non-aberrant transcripts. In animals, the SMG1 kinase plays an essential role in NMD by phosphorylating the core NMD factor UPF1. Despite SMG1 being ubiquitous throughout the plant kingdom, little is known about its function, probably because SMG1 is atypically absent from the genome of the model plant, Arabidopsis thaliana. By combining our previously established SMG1 knockout in moss with transcriptome-wide analysis, we reveal the range of processes involving SMG1 in plants. Machine learning assisted analysis suggests that 32% of multi-isoform genes produce NMD-targeted transcripts and that splice junctions downstream of a stop codon act as the major determinant of NMD targeting. Furthermore, we suggest that SMG1 is involved in other quality control pathways, affecting DNA repair and the unfolded protein response, in addition to its role in mRNA quality control. Consistent with this, smg1 plants have increased susceptibility to DNA damage, but increased tolerance to unfolded protein inducing agents. The potential involvement of SMG1 in RNA, DNA and protein quality control has major implications for the study of these processes in plants.
Asunto(s)
Bryopsida/enzimología , Bryopsida/genética , Degradación de ARNm Mediada por Codón sin Sentido , Fosfotransferasas/fisiología , Proteínas de Plantas/fisiología , Regiones no Traducidas 3' , Bryopsida/metabolismo , Daño del ADN , Expresión Génica , Mutación , Fosfotransferasas/genética , Proteínas de Plantas/genética , Respuesta de Proteína DesplegadaRESUMEN
Nonsense-mediated mRNA decay (NMD) is an essential eukaryotic process regulating transcript quality and abundance, and is involved in diverse processes including brain development and plant defenses. Although some of the NMD machinery is conserved between kingdoms, little is known about its evolution. Phosphorylation of the core NMD component UPF1 is critical for NMD and is regulated in mammals by the SURF complex (UPF1, SMG1 kinase, SMG8, SMG9 and eukaryotic release factors). However, since SMG1 is reportedly missing from the genomes of fungi and the plant Arabidopsis thaliana, it remains unclear how UPF1 is activated outside the metazoa. We used comparative genomics to determine the conservation of the NMD pathway across eukaryotic evolution. We show that SURF components are present in all major eukaryotic lineages, including fungi, suggesting that in addition to UPF1 and SMG1, SMG8 and SMG9 also existed in the last eukaryotic common ancestor, 1.8 billion years ago. However, despite the ancient origins of the SURF complex, we also found that SURF factors have been independently lost across the Eukarya, pointing to genetic buffering within the essential NMD pathway. We infer an ancient role for SURF in regulating UPF1, and the intriguing possibility of undiscovered NMD regulatory pathways.
Asunto(s)
Eucariontes/genética , Evolución Molecular , Complejos Multienzimáticos/genética , Degradación de ARNm Mediada por Codón sin Sentido/genética , Genómica/métodosRESUMEN
Gene amplification followed by functional diversification is a major force in evolution. A typical example of this is seen in the WUSCHEL-RELATED HOMEOBOX (WOX) gene family, named after the Arabidopsis stem cell regulator WUSCHEL. Here we analyze functional divergence in the WOX gene family. Members of the WUS clade, except the cambium stem cell regulator WOX4, can substitute for WUS function in shoot and floral stem cell maintenance to different degrees. Stem cell function of WUS requires a canonical WUS-box, essential for interaction with TPL/TPR co-repressors, whereas the repressive EAR domain is dispensable and the acidic domain seems only to be required for female fertility. In contrast to the WUS clade, members of the ancient WOX13 and the WOX9 clades cannot support stem cell maintenance. Although the homeodomains are interchangeable between WUS and WOX9 clade members, a WUS-compatible homeodomain together with canonical WUS-box is not sufficient for stem cell maintenance. Our results suggest that WOX function in shoot and floral meristems of Arabidopsis is restricted to the modern WUS clade, suggesting that stem cell control is a derived function. Yet undiscovered functional domains in addition to the homeodomain and the WUS-box are necessary for this function.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/citología , Arabidopsis/metabolismo , Células Madre/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Meristema/citología , Meristema/metabolismo , Brotes de la Planta/citología , Brotes de la Planta/metabolismo , Células Madre/fisiologíaRESUMEN
A complete understanding of the genetic control of flower development requires a comparative approach, involving species from across the angiosperm lineage. Using the accessible model plant Arabidopsis thaliana many of the genetic pathways that control development of the reproductive growth phase have been delineated. Research in other species has added to this knowledge base, revealing that, despite the myriad of floral forms found in nature, the genetic blueprint of flower development is largely conserved. However, these same studies have also highlighted differences in the way flowering is controlled in evolutionarily diverse species. Here, we review flower development in the eudicot asterid lineage, a group of plants that diverged from the rosid family, which includes Arabidopsis, 120 million years ago. Work on model species such as Antirrhinum majus, Petunia hybrida, and Gerbera hybrida has prompted a reexamination of textbook models of flower development; revealed novel mechanisms controlling floral gene expression; provided a means to trace evolution of key regulatory genes; and stimulated discussion about genetic redundancy and the fate of duplicated genes.
Asunto(s)
Flores/crecimiento & desarrollo , Magnoliopsida/crecimiento & desarrollo , Flores/metabolismo , Magnoliopsida/metabolismo , Meristema/crecimiento & desarrollo , Meristema/metabolismoRESUMEN
Large-scale protein-protein interaction studies recently demonstrated that the Arabidopsis TPL/TPR family of transcriptional co-repressors is involved in a broad range of developmental processes. TPL/TPRs predominantly interact with transcription factors that contain repression domain (RD) sequences. Interestingly, RDs reported in the literature are quite diverse in sequence, yet TPL/TPRs interact with proteins containing all of the known motifs. These data lead us to conclude that the TPL/TPRs act as general repressors of gene transcription in plants. To investigate this further, we examined interactions between TPL/TPR proteins encoded by the moss Physcomitrella patens genome and components of the auxin signaling pathway. As in Arabidopsis, moss TPL proteins interact with AUX/IAA and ARF proteins, suggesting that they act in both forms of ARF-mediated transcriptional repression. These data suggest that the involvement of TPL in auxin signaling has been conserved across evolution, since mosses and angiosperms diverged approximately 450 million years ago.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas Co-Represoras/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/clasificación , Proteínas de Arabidopsis/genética , Proteínas Co-Represoras/clasificación , Proteínas Co-Represoras/genética , Evolución Molecular , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Genoma de Planta/genética , Filogenia , Unión Proteica/genética , Unión Proteica/fisiología , Transducción de Señal/genética , Transducción de Señal/fisiologíaRESUMEN
Transcription factors activate or repress target gene expression or switch between activation and repression. In animals and yeast, Groucho/Tup1 corepressor proteins are recruited by diverse transcription factors to induce context-specific transcriptional repression. Two groups of Groucho/Tup1-like corepressors have been described in plants. LEUNIG and LEUNIG_HOMOLOG constitute one group and TOPLESS (TPL) and the four TPL-related (TPR) corepressors form the other. To discover the processes in which TPL and the TPR corepressors operate, high-throughput yeast two-hybrid approaches were used to identify interacting proteins. We found that TPL/TPR corepressors predominantly interact directly with specific transcription factors, many of which were previously implicated in transcriptional repression. The interacting transcription factors reveal that the TPL/TPR family has been coopted multiple times to modulate gene expression in diverse processes, including hormone signaling, stress responses, and the control of flowering time, for which we also show biological validation. The interaction data suggest novel mechanisms for the involvement of TPL/TPR corepressors in auxin and jasmonic acid signaling. A number of short repression domain (RD) sequences have previously been identified in Arabidopsis (Arabidopsis thaliana) transcription factors. All known RD sequences were enriched among the TPL/TPR interactors, and novel TPL-RD interactions were identified. We show that the presence of RD sequences is essential for TPL/TPR recruitment. These data provide a framework for TPL/TPR-dependent transcriptional repression. They allow for predictions about new repressive transcription factors, corepressor interactions, and repression mechanisms and identify a wide range of plant processes that utilize TPL/TPR-mediated gene repression.
Asunto(s)
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Arabidopsis/metabolismo , Secuencia de Bases , Flores/fisiología , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Datos de Secuencia Molecular , Proteínas de Complejo Poro Nuclear/genética , Proteínas de Complejo Poro Nuclear/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Plantas Modificadas Genéticamente , Mapas de Interacción de Proteínas , Estructura Terciaria de Proteína , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Técnicas del Sistema de Dos HíbridosRESUMEN
The evolution of the floral homeotic genes has been characterized using phylogenetic and functional studies. It is possible to enhance these studies by comparing gene content and order between species to determine the evolutionary history of the regulatory genes. Here, we use a synteny-based approach to trace the evolution of the floral B- and C-function genes that are required for specification of the reproductive organs. Consistent with previous phylogenetic studies, we show that the euAP3-TM6 split occurred after the monocots and dicots diverged. The Arabidopsis TM6 and papaya euAP3 genes are absent from the respective genomes, and we have detected loci from which these genes were lost. These data indicate that either the TM6 or the euAP3 lineage genes can be lost without detriment to flower development. In contrast, PI is essential for male reproductive organ development; yet, contrary to predictions, complex genomic rearrangements have resulted in almost complete breakdown of synteny at the PI locus. In addition to showing the evolution of B-function genes through the prediction of ancestral loci, similar reconstructions reveal the origins of the C-function AG and PLE lineages in dicots, and show the shared ancestry with the monocot C-function genes. During our studies, we found that transposable elements (TEs) present in sequenced Antirrhinum genomic clones limited comparative studies. A pilot survey of the Antirrhinum data revealed that gene-rich regions contain an unusually high degree of TEs of very varied types, which will be an important consideration for future genome sequencing efforts.
Asunto(s)
Antirrhinum/genética , Evolución Molecular , Flores/genética , Genes Homeobox/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Sintenía/genética , Cromosomas Artificiales Bacterianos/genética , Secuencia Conservada/genética , Sitios Genéticos/genética , FilogeniaRESUMEN
One of the early successes of the application of molecular genetics to study plant development was the discovery of a series of genes that act together, in an apparently simple combinatorial model, to specify the identity of the different organs of a flower. Widely known as the ABC model, this framework for understanding has been investigated and modified over the course of the last two decades. The cast list of genes has been defined and, as other chapters in this volume will show, great progress has been made in understanding how they are regulated, how they act together, what they do and how they have contributed to the evolution of the flower in its varied forms. In this introductory review to the volume we will review the derivation and elaboration of the most current version of the ABC model, highlighting the modifications that have been necessary to ensure it fits the available experimental data. We will highlight the remaining difficulties in fitting the current model to the experimental data and propose a further modification to enable it to regain its applicability.
Asunto(s)
Flores , Modelos Biológicos , Arabidopsis/anatomía & histología , Arabidopsis/crecimiento & desarrollo , Arabidopsis/fisiología , Evolución Biológica , Flores/anatomía & histología , Flores/crecimiento & desarrollo , Flores/fisiología , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
Control of organ size is the product of coordinated cell division and expansion. In plants where one of these pathways is perturbed, organ size is often unaffected as compensation mechanisms are brought into play. The number of founder cells in organ primordia, dividing cells, and the period of cell proliferation determine cell number in lateral organs. We have identified the Antirrhinum FORMOSA (FO) gene as a specific regulator of floral size. Analysis of cell size and number in the fo mutant, which has increased flower size, indicates that FO is an organ-specific inhibitor of cell division and activator of cell expansion. Increased cell number in fo floral organs correlated with upregulation of genes involved in the cell cycle. In Arabidopsis the AINTEGUMENTA (ANT) gene promotes cell division. In the fo mutant increased cell number also correlates with upregulation of an Antirrhinum ANT-like gene (Am-ANT) in inflorescences that is very closely related to ANT and shares a similar expression pattern, suggesting that they may be functional equivalents. Increased cell proliferation is thought to be compensated for by reduced cell expansion to maintain organ size. In Arabidopsis petal cell expansion is inhibited by the BIGPETAL (BPE) gene, and in the fo mutant reduced cell size corresponded to upregulation of an Antirrhinum BPE-like gene (Am-BPE). Our data suggest that FO inhibits cell proliferation by negatively regulating Am-ANT, and acts upstream of Am-BPE to coordinate floral organ size. This demonstrates that organ size is modulated by the organ-specific control of both general and local gene networks.
Asunto(s)
Antirrhinum/genética , Flores/genética , Proteínas de Plantas/genética , Secuencia de Aminoácidos , Antirrhinum/citología , Antirrhinum/crecimiento & desarrollo , Ciclo Celular , División Celular , Tamaño de la Célula , Ciclina D , Ciclinas/genética , Flores/citología , Flores/ultraestructura , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Hibridación in Situ , Microscopía Electrónica de Rastreo , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/clasificación , Proteínas de Plantas/fisiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de AminoácidoRESUMEN
The floral C-function, which specifies stamen and carpel development, played a pivotal role in the evolution of flowers. An important aspect of this was the establishment of mechanisms regulating the temporal and spatial expression domain of the C-function genes. Transcription of the Arabidopsis C-function gene AGAMOUS (AG) is tightly controlled by factors that interact with cis-elements within its large second intron. Little is known about the regulatory role of intragenic elements in C-function genes from species other than Arabidopsis. We show that a binding site for the LEAFY (LFY) transcription factor, present in the AG intron, is conserved in the introns of diverse C-function genes and is positioned close to other conserved motifs. Using an in planta mutagenesis approach, we targeted evolutionarily conserved sequences in the intron of the Antirrhinum PLENA (PLE) gene to establish whether they regulate PLE expression. Small sequence deletions resulted in a novel class of heterochronic C-function mutants with delayed onset of PLE expression and loss of stamen identity. These phenotypes differ significantly from weak C-function mutant alleles in Antirrhinum and Arabidopsis. Our findings demonstrate that the PLE intron contains regulatory cis-elements, including a LFY-binding site, critical for establishing the correct C-function expression domain. We show that the LFY site, and other conserved intron elements, pre-date the divergence of the monocot and dicot lineages, suggesting that they were a determinant in the evolution of the C-function, and propose a threshold model to explain phenotypic divergence observed between C-function mutants.
Asunto(s)
Antirrhinum/genética , Evolución Molecular , Flores/genética , Proteínas de Dominio MADS/genética , Proteínas de Plantas/genética , Alelos , Antirrhinum/clasificación , Antirrhinum/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Secuencia de Bases , Sitios de Unión , Secuencia Conservada , Flores/metabolismo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Intrones , Proteínas de Dominio MADS/clasificación , Proteínas de Dominio MADS/metabolismo , Fenotipo , Filogenia , Proteínas de Plantas/clasificación , Proteínas de Plantas/metabolismo , Elementos Reguladores de la Transcripción , Eliminación de Secuencia , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Gene duplication plays a fundamental role in evolution by providing the genetic material from which novel functions can arise. Newly duplicated genes can be maintained by subfunctionalization (the duplicated genes perform different aspects of the original gene's function) and/or neofunctionalization (one of the genes acquires a novel function). PLENA in Antirrhinum and AGAMOUS in Arabidopsis are the canonical C-function genes that are essential for the specification of reproductive organs. These functionally equivalent genes encode closely related homeotic MADS-box transcription factors. Using genome synteny, we confirm phylogenetic analyses showing that PLENA and AGAMOUS are nonorthologous genes derived from a duplication in a common ancestor. Their respective orthologs, SHATTERPROOF in Arabidopsis and FARINELLI in Antirrhinum, have undergone independent subfunctionalization via changes in regulation and protein function. Surprisingly, the functional divergence between PLENA and FARINELLI, is morphologically manifest in both transgenic Antirrhinum and Arabidopsis. This provides a clear illustration of a random evolutionary trajectory for gene functions after a duplication event. Different members of a duplicated gene pair have retained the primary homeotic functions in different lineages, illustrating the role of chance in evolution. The differential ability of the Antirrhinum genes to promote male or female development provides a striking example of subfunctionalization at the protein level.
Asunto(s)
Proteína AGAMOUS de Arabidopsis/fisiología , Antirrhinum/genética , Arabidopsis/genética , Evolución Molecular , Flores/genética , Genes Duplicados/fisiología , Proteínas de Dominio MADS/fisiología , Proteínas de Plantas/fisiología , Proteína AGAMOUS de Arabidopsis/genética , Proteína AGAMOUS de Arabidopsis/metabolismo , Antirrhinum/fisiología , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/fisiología , Secuencia de Bases , Cromosomas Artificiales Bacterianos , Biología Computacional , Flores/fisiología , Componentes del Gen , Genes Duplicados/genética , Vectores Genéticos , Proteínas de Dominio MADS/genética , Proteínas de Dominio MADS/metabolismo , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Rhizobium , Análisis de Secuencia de ADN , Sintenía/genética , Factores de Transcripción/genética , Factores de Transcripción/fisiologíaRESUMEN
Protein interactions are crucial to the life of a cell. The analysis of such interactions is allowing biologists to determine the function of uncharacterized proteins and the genes that encode them. The yeast two-hybrid system has become one of the most popular and powerful tools to study protein-protein interactions. With the advent of proteomics, the two-hybrid system has found a niche in interactome mapping. However, it is clear that only by combining two-hybrid data with that from complementary approaches such as mass spectrometry (MS) can the interactome be analyzed in full. This review introduces the yeast two-hybrid system to those unfamiliar with the technique, and discusses how it can be used in combination with MS to unravel the network of protein interactions that occur in a cell.
Asunto(s)
Espectrometría de Masas/métodos , Proteínas/genética , Proteómica/instrumentación , Levaduras/genética , Genes Fúngicos/genética , Genoma , Proteínas/química , Proteínas/fisiología , Proteómica/métodos , Levaduras/fisiologíaRESUMEN
Cupuliformis mutants are defective in shoot apical meristem formation, but cup plants overcome this early barrier to development to reach maturity. CUP encodes a NAC-domain transcription factor, homologous to the Petunia NAM and Arabidopsis CUC proteins. The phenotype of cup mutants differs from those of nam and cuc1 cuc2 in that dramatic organ fusion is observed throughout development. In addition to cotyledon and floral organ fusions, severe lateral organ fusion is found in leaves and inflorescences, and the apical meristem becomes highly fasciated. These features reveal a role for CUP in the establishment of all above ground organ boundaries. Consistent with this function, CUP is expressed at the boundaries of all lateral organs and meristems. It is not currently known how NAC-domain genes act to establish organ boundaries. Here, we show that CUP directly interacts with a TCP-domain transcription factor. Members of the TCP-domain family have previously been shown to regulate organ outgrowth. Our results suggest a model for the establishment of organ boundaries based on the localised expression of NAC-domain and TCP-domain factors.
Asunto(s)
Antirrhinum/embriología , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The development of floral reproductive organs requires the activity of plant MADS-box transcription factors (MBFs) belonging to the C function. The C function can only operate within a floral context, specified by MBFs belonging to the SEPALLATA class of proteins. Here we describe the specific interaction between a novel protein, MIP1, and C-function and SEPALLATA (SEP)-like MBFs. MIP1 is the first member of a new class of proteins unique to plants. None of the family members have yet been assigned a function. Motif searches reveal a leucine zipper domain within a conserved N-terminal region of MIP1. The leucine zipper lies within a region sufficient for interaction with plant MBFs. MIP1 interacts with a domain of plant MBFs that is analogous to the domain of animal and yeast MBFs involved in ternary complex formation. The MIP1 protein is predicted to localise to the nucleus and activates yeast reporter genes in vivo. MIP1 is expressed in the fourth whorl of the flower, in an overlapping temporal and spatial expression pattern with the C-function and SEP-like genes. Taken together, this suggests that MIP1 acts as a ternary complex factor specifically with C-function and SEP-like MBFs.
Asunto(s)
Antirrhinum/genética , Proteínas Portadoras/genética , Proteínas de Dominio MADS/metabolismo , Proteínas de Plantas/metabolismo , Secuencia de Aminoácidos , Antirrhinum/metabolismo , Secuencia de Bases , Sitios de Unión/genética , Proteínas Portadoras/metabolismo , ADN Complementario/química , ADN Complementario/genética , ADN Complementario/aislamiento & purificación , Flores/genética , Regulación de la Expresión Génica de las Plantas , Hibridación in Situ , Proteínas de Dominio MADS/genética , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/genética , Unión Proteica , Saccharomyces cerevisiae/genética , Análisis de Secuencia de ADN , Técnicas del Sistema de Dos HíbridosAsunto(s)
Frutas/fisiología , Genes de Plantas , Proteínas de Dominio MADS/genética , Proteínas de Plantas/genética , Solanum lycopersicum/fisiología , Factores de Transcripción/genética , Etilenos/biosíntesis , Etilenos/metabolismo , Regulación de la Expresión Génica de las Plantas , Solanum lycopersicum/genética , Proteínas de Dominio MADS/fisiología , Mutación , Proteínas de Plantas/fisiología , Eliminación de Secuencia , Factores de Transcripción/fisiologíaRESUMEN
Plant research is moving into the post-genomic era. Proteomic-based strategies are now being developed to study functional aspects of the genes predicted from the various genome-sequencing initiatives. All biological processes depend on interactions formed between proteins and the mapping of such interactions on a global scale is providing interesting functional insights. One of the techniques that has proved itself invaluable in the mapping of protein-protein interactions is the yeast two-hybrid system. This system is a sensitive molecular genetic approach for studying protein-protein interactions in vivo. In this review we will introduce the yeast two-hybrid system, discuss modifications of the system that may be of interest to the plant science community and suggest potential applications of the technology.