ABSTRACT
The basic helix-loop-helix transcription factor (bHLH TF) family is involved in tissue development, cell differentiation, and disease. These factors have transcriptionally positive, negative, and inactive functions by combining dimeric interactions among family members. The best known bHLH TFs are the E-protein homodimers and heterodimers with the tissue-specific TFs or ID proteins. These cooperative and dynamic interactions result in a complex transcriptional network that helps define the cell's fate. Here, the reported dimeric interactions of 67 vertebrate bHLH TFs with other family members are summarized in tables, including specifications of the experimental techniques that defined the dimers. The compilation of these extensive data underscores homodimers of tissue-specific bHLH TFs as a central part of the bHLH regulatory network, with relevant positive and negative transcriptional regulatory roles. Furthermore, some sequence-specific TFs can also form transcriptionally inactive heterodimers with each other. The function, classification, and developmental role for all vertebrate bHLH TFs in four major classes are detailed.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/chemistry , Basic Helix-Loop-Helix Transcription Factors/metabolism , Dimerization , Protein Multimerization , Animals , Basic Helix-Loop-Helix Transcription Factors/classification , Humans , Models, Chemical , Protein Structure, QuaternaryABSTRACT
The phytochrome-interacting factors (PIFs) proteins belong to the subfamily of basic helix-loop-helix (bHLH) transcription factors and play important roles in chloroplast development and chlorophyll biosynthesis. Currently, knowledge about the PIF gene family in Camellia sinensis remains very limited. In this study, seven PIF members were identified in the C. sinensis genome and named based on homology with AtPIF genes in Arabidopsis thaliana. All C. sinensis PIF (CsPIF) proteins have both the conserved active PHYB binding (APB) and bHLH domains. Phylogenetic analysis revealed that CsPIFs were clustered into four groups-PIF1, PIF3, PIF7, and PIF8-and most CsPIFs were clustered in pairs with their corresponding orthologs in Populus tremula. CsPIF members in the same group tended to display uniform or similar exon-intron distribution patterns and motif compositions. CsPIF genes were differentially expressed in C. sinensis with various leaf colors and strongly correlated with the expression of genes involved in the chlorophyll metabolism pathway. Promoter analysis of structural genes related to chlorophyll metabolism found DNA-binding sites of PIFs were abundant in the promoter regions. Protein-protein interaction networks of CsPIFs demonstrated a close association with phytochrome, PIF4, HY5, TOC1, COP1, and PTAC12 proteins. Additionally, subcellular localization and transcriptional activity analysis suggested that CsPIF3b was nuclear localized protein and possessed transcriptional activity. We also found that CsPIF3b could activate the transcription of CsHEMA and CsPOR in Nicotiana benthamiana leaves. This work provides comprehensive research of CsPIFs and would be helpful to further promote the regulation mechanism of PIF on chlorophyll metabolism in C. sinensis.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/metabolism , Camellia sinensis/metabolism , Chlorophyll/metabolism , Plant Proteins/metabolism , Amino Acid Sequence , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/genetics , Gene Expression Regulation, Plant , Phylogeny , Plant Leaves/metabolism , Plant Proteins/classification , Plant Proteins/genetics , Protein Interaction Maps/genetics , Protein Isoforms/classification , Protein Isoforms/genetics , Protein Isoforms/metabolism , Sequence Alignment , Transcriptional ActivationABSTRACT
Physical interactions between members of the MYB and bHLH transcription factor (TF) families regulate many important biological processes in plants. Not all reported MYB-bHLH interactions can be explained by the known binding sites in the R3 repeat of the MYB DNA-binding domain. Noteworthy, most of the sequence diversity of MYB TFs lies in their non-MYB regions, which contain orphan small subgroup-defining motifs not yet linked to molecular functions. Here, we identified the motif mediating interaction between MYB TFs from subgroup 12 and their bHLH partners. Unlike other known MYB-bHLH interactions, the motif locates to the centre of the predicted disordered non-MYB region. We characterised the core motif, which enabled accurate prediction of previously unknown bHLH-interacting MYB TFs in Arabidopsis thaliana, and we confirmed its functional importance in planta. Our results indicate a correlation between the MYB-bHLH interaction affinity and the phenotypic output controlled by the TF complex. The identification of an interaction motif outside R3 indicates that MYB-bHLH interactions must have arisen multiple times, independently and suggests many more motifs of functional relevance to be harvested from subgroup-specific studies.
Subject(s)
Arabidopsis Proteins/genetics , Basic Helix-Loop-Helix Transcription Factors/genetics , DNA-Binding Proteins/genetics , Phylogeny , Transcription Factors/genetics , Amino Acid Sequence/genetics , Arabidopsis/genetics , Arabidopsis Proteins/classification , Basic Helix-Loop-Helix Transcription Factors/classification , Gene Expression Regulation, Plant/genetics , Plants, Genetically Modified/genetics , Transcription Factors/classificationABSTRACT
Teosinte branched 1/Cycloidea/Proliferating cell factor 1 (TCP) proteins belongs to a plant-specific transcription factor family that plays important roles in plant development. TCP gene-regulated plant branching occurs downstream in the strigolactone pathway. In this study, 41 TCP genes were identified in the genome of Panicum virgatum L. (switchgrass). These genes all contained the TCP conserved domain, and they belonged to two subfamilies distributed across 18 chromosomes. Analysis of gene expression using RNA-Seq data showed that 16 TCP genes were highly expressed in the inflorescence and shoot. The expression patterns of 13 selected PvTCP genes were analyzed in different tissues, and their responses to strigolactones (SLs) were examined. The selected genes were expressed differentially in a range of tissues and to application of SLs, indicating that PvTCPs were involved in a range of developmental and physiological processes. This genome-wide analysis and determination of PvTCP gene-expression patterns yielded valuable information on switchgrass development that will inform studies into improving switchgrass and other species for crop production.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Multigene Family , Panicum/genetics , Plant Proteins/genetics , Basic Helix-Loop-Helix Transcription Factors/chemistry , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/metabolism , Chromosomes, Plant , Genes, Plant , Panicum/metabolism , Plant Growth Regulators/pharmacology , Plant Proteins/chemistry , Plant Proteins/classification , Plant Proteins/metabolism , SyntenyABSTRACT
Paclitaxel is an important anticancer drug. The phytohormone jasmonic acid can significantly induce the biosynthesis of paclitaxel in Taxus, but the molecular mechanism has not yet been resolved. To establish the jasmonic acid signalling pathway of Taxus media, based on the gene of the jasmonic acid signalling pathway of Arabidopsis thaliana, sequence analysis was performed to isolate the jasmonic acid signal from the transcriptome, a transcriptional cluster of pathway gene homologs and the full length of 22 genes were obtained by RACE PCR at 5' and 3': two EI ubiquitin ligase genes, COI1-1 and COI1-2;7 MYC bHLH type transcription factor (MYC2, MYC3, MYC4, JAM1, JAM2, EGL3, TT8); 12 JAZ genes containing the ZIM domain; and MED25, one of the components of the transcriptional complex. The protein interaction between each were confirmed by yeast two hybridization and bimolecular fluorescence complementation based on similar genes interaction in Arabidopsis. A similar jasmonate signaling pathway was illustrated in T. media. All known paclitaxel biosynthesis genes promoters were isolated by genome walker PCR. To investigate the jasmonate signaling effect on these genes' expression, the transcription activity of MYC2, MYC3 and MYC4 on these promoters were examined. There are 12, 10 and 11 paclitaxel biosynthesis genes promoters that could be activated by MYC2, MYC3 and MYC4.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/metabolism , Cyclopentanes/metabolism , Oxylipins/metabolism , Paclitaxel/biosynthesis , Plant Proteins/metabolism , Taxus/metabolism , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/genetics , Phylogeny , Plant Proteins/classification , Plant Proteins/genetics , Promoter Regions, Genetic , Signal Transduction , Two-Hybrid System TechniquesABSTRACT
The tea plant is an important commercial horticulture crop cultivated worldwide. Yield and quality of this plant are influenced by abiotic stress. The bHLH family transcription factors play a pivotal role in the growth and development, including abiotic stress response, of plants. A growing number of bHLH proteins have been functionally characterized in plants. However, few studies have focused on the bHLH proteins in tea plants. In this study, 120 CsbHLH TFs were identified from tea plants using computational prediction method. Structural analysis detected 23 conservative residues, with over 50% identities in the bHLH domain. Moreover, 103 CsbHLH proteins were assumed to bind DNA and encompassed 98 E-Box binders and 85 G-Box binders. The CsbHLH proteins were grouped into 20 subfamilies based on phylogenetic analysis and a previous classification system. A survey of transcriptome profiling screened 22 and 39 CsbHLH genes that were upregulated under heat and drought stress. Nine CsbHLH genes were validated using qRT-PCR. Results were approximately in accordance with transcriptome data. These genes could be induced by one or more abiotic stresses.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Camellia sinensis/genetics , DNA, Plant/genetics , Gene Expression Regulation, Plant , Plant Proteins/genetics , Transcriptome , Adaptation, Physiological/genetics , Amino Acid Sequence , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/metabolism , Camellia sinensis/classification , Camellia sinensis/metabolism , Conserved Sequence , DNA, Plant/metabolism , Droughts , Hot Temperature , Multigene Family , Phylogeny , Plant Proteins/classification , Plant Proteins/metabolism , Protein Binding , Sequence Alignment , Stress, PhysiologicalABSTRACT
Plants have evolved different abilities to adapt to the ever-fluctuating environments for sessility. Calcium-dependent protein kinase (CDPK) is believed to play a pivotal role in abiotic stress signaling. So far, study on the specific substrates that CDPK recognized in response to adversity is limited. In the present study, we revealed a potential interaction between CDPK and a bHLH transcription factor under salt stress in Chenopodium glaucum. First, we identified a CgCDPK, which was up-regulated under salt and drought stress; then by Y2H screening, CgCDPK was detected to be involved in interaction with a bHLH TF (named as CgbHLH001), which also positively respond to salt and drought stress. Further computational prediction and experiments including GST-pulldown and BiFC assays revealed that potential interaction existed between CgCDPK and CgbHLH001, and they might interact on the plasma membrane. In addition, CgCDPK-overexpressed transgenic tobacco line could significantly accumulate transcripts of NtbHLH (a homolog of CgbHLH001 in N. tabacum), which provided another evidence of correlation between CgCDPK and CgbHLH001. Our results suggest that CgbHLH001 can interact with CgCDPK in signal transduction pathway in response to abiotic stress, which should provide new evidence for further understanding of the substrate specificity of plant CDPK signaling pathway.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/metabolism , Chenopodium/metabolism , Plant Proteins/metabolism , Protein Kinases/metabolism , Amino Acid Sequence , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/genetics , Cell Membrane/metabolism , Chenopodium/genetics , Droughts , Gene Expression Profiling , Gene Expression Regulation, Plant/drug effects , Phylogeny , Plant Proteins/classification , Plant Proteins/genetics , Protein Binding , Protein Kinases/classification , Protein Kinases/genetics , Sequence Homology, Amino Acid , Sodium Chloride/pharmacology , Stress, Physiological , Two-Hybrid System TechniquesABSTRACT
Hypoxia inducible factor (HIF) transcription factors are crucial for regulating a variety of cellular activities in response to oxygen stress (hypoxia). In this study, we determine the evolutionary history of HIF genes and their associated transactivation domains, as well as perform selection and functional divergence analyses across their four characteristic domains. Here we show that the HIF genes are restricted to metazoans: At least one HIF-α homolog is found within the genomes of non-bilaterians and bilaterian invertebrates, while most vertebrate genomes contain between two and six HIF-α genes. We also find widespread purifying selection across all four characteristic domain types, bHLH, PAS, NTAD, CTAD, in HIF-α genes, and evidence for Type I functional divergence between HIF-1α, HIF-2α /EPAS, and invertebrate HIF genes. Overall, we describe the evolutionary histories of the HIF transcription factor gene family and its associated transactivation domains in eukaryotes. We show that the NTAD and CTAD domains appear de novo, without any appearance outside of the HIF-α subunits. Although they both appear in invertebrates as well as vertebrate HIF- α sequences, there seems to have been a substantial loss across invertebrates or were convergently acquired in these few lineages. We reaffirm that HIF-1α is phylogenetically conserved among most metazoans, whereas HIF-2α appeared later. Overall, our findings can be attributed to the substantial integration of this transcription factor family into the critical tasks associated with maintenance of oxygen homeostasis and vascularization, particularly in the vertebrate lineage.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Eukaryota/genetics , Genetic Variation , Selection, Genetic , Vertebrates/genetics , Animals , Basic Helix-Loop-Helix Transcription Factors/classification , Eukaryota/classification , Evolution, Molecular , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/classification , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Phylogeny , Vertebrates/classificationABSTRACT
The basic helix-loop-helix (bHLH) transcription factors (TFs) form a huge superfamily and play crucial roles in many essential developmental, genetic, and physiological-biochemical processes of eukaryotes. In total, 109 putative bHLH TFs were identified and categorized successfully in the genomic databases of cattle, Bos Taurus, after removing redundant sequences and merging genetic isoforms. Through phylogenetic analyses, 105 proteins among these bHLH TFs were classified into 44 families with 46, 25, 14, 3, 13, and 4 members in the high-order groups A, B, C, D, E, and F, respectively. The remaining 4 bHLH proteins were sorted out as 'orphans.' Next, these 109 putative bHLH proteins identified were further characterized as significantly enriched in 524 significant Gene Ontology (GO) annotations (corrected P value ≤ 0.05) and 21 significantly enriched pathways (corrected P value ≤ 0.05) that had been mapped by the web server KOBAS 2.0. Furthermore, 95 bHLH proteins were further screened and analyzed together with two uncharacterized proteins in the STRING online database to reconstruct the protein-protein interaction network of cattle bHLH TFs. Ultimately, 89 bHLH proteins were fully mapped in a network with 67 biological process, 13 molecular functions, 5 KEGG pathways, 12 PFAM protein domains, and 25 INTERPRO classified protein domains and features. These results provide much useful information and a good reference for further functional investigations and updated researches on cattle bHLH TFs.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Basic Helix-Loop-Helix Transcription Factors/isolation & purification , Genome/genetics , Amino Acid Sequence/genetics , Animals , Basic Helix-Loop-Helix Transcription Factors/classification , Cattle , Molecular Sequence Annotation , PhylogenyABSTRACT
ROOT HAIR SPECIFIC (RHS) genes, which contain the root hair-specific cis-element (RHE) in their regulatory regions, function in root hair morphogenesis. Here, we demonstrate that an Arabidopsis thaliana basic helix-loop-helix transcription factor, ROOT HAIR DEFECTVE SIX-LIKE4 (RSL4), directly binds to the RHE in vitro and in vivo, upregulates RHS genes, and stimulates root hair formation in Arabidopsis. Orthologs of RSL4 from a eudicot (poplar [Populus trichocarpa]), a monocot (rice [Oryza sativa]), and a lycophyte (Selaginella moellendorffii) each restored root hair growth in the Arabidopsis rsl4 mutant. In addition, the rice and S. moellendorffii RSL4 orthologs bound to the RHE in in vitro and in vivo assays. The RSL4 orthologous genes contain RHEs in their promoter regions, and RSL4 was able to bind to its own RHEs in vivo and amplify its own expression. This process likely provides a positive feedback loop for sustainable root hair growth. When RSL4 and its orthologs were expressed in cells in non-root-hair positions, they induced ectopic root hair growth, indicating that these genes are sufficient to specify root hair formation. Our results suggest that RSL4 mediates root hair formation by regulating RHS genes and that this mechanism is conserved throughout the tracheophyte (vascular plant) lineage.
Subject(s)
Arabidopsis Proteins/genetics , Basic Helix-Loop-Helix Transcription Factors/genetics , Gene Expression Regulation, Plant , Plant Roots/genetics , Plant Structures/genetics , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/classification , Arabidopsis Proteins/metabolism , Base Sequence , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/metabolism , Microscopy, Confocal , Oryza/genetics , Oryza/metabolism , Phylogeny , Plant Roots/metabolism , Plant Structures/metabolism , Plants, Genetically Modified , Populus/genetics , Populus/metabolism , Promoter Regions, Genetic/genetics , Reverse Transcriptase Polymerase Chain Reaction , Selaginellaceae/genetics , Selaginellaceae/metabolism , Sequence Homology, Nucleic AcidABSTRACT
Transcription factors of the basic helix-loop-helix (bHLH) PER-ARNT-SIM (PAS) family generally have critical and nonredundant biological roles, but some bHLH PAS proteins compete for common cofactors or recognise similar DNA elements. Identifying factors that regulate function of bHLH PAS proteins, particularly in cells where multiple family members are coexpressed, is important for understanding bHLH PAS factor biology. This study identifies and characterises a novel interaction between melanoma-associated antigen D1 (MAGED1) and select members of the bHLH PAS transcription factor family. MAGED1 binds and positively regulates the transcriptional activity of family members SIM1, SIM2, NPAS4 and ARNT2, but does not interact with AhR, HIF1α and ARNT. This interaction is mediated by PAS repeat regions which also form the interface for bHLH PAS dimerisation, and accordingly MAGED1 is not found in complex with bHLH PAS dimers. We show that MAGED1 does not affect bHLH PAS protein levels and cannot be acting as a coactivator of transcriptionally active heterodimers, but rather appears to interact with nascent bHLH PAS proteins in the cytoplasm to enhance their function prior to nuclear import. As a selective regulator, MAGED1 may play an important role in the biology of these specific factors and in general bHLH PAS protein dynamics.
Subject(s)
Antigens, Neoplasm/metabolism , Basic Helix-Loop-Helix Transcription Factors/metabolism , Neoplasm Proteins/metabolism , Protein Serine-Threonine Kinases/metabolism , Antigens, Neoplasm/chemistry , Antigens, Neoplasm/genetics , Aryl Hydrocarbon Receptor Nuclear Translocator/chemistry , Aryl Hydrocarbon Receptor Nuclear Translocator/genetics , Aryl Hydrocarbon Receptor Nuclear Translocator/metabolism , Basic Helix-Loop-Helix Transcription Factors/chemistry , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/genetics , HEK293 Cells , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/chemistry , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Neoplasm Proteins/chemistry , Neoplasm Proteins/genetics , Protein Interaction Domains and Motifs , Protein Isoforms/chemistry , Protein Isoforms/genetics , Protein Isoforms/metabolism , Protein Serine-Threonine Kinases/classification , Protein Stability , Receptors, Aryl Hydrocarbon/chemistry , Receptors, Aryl Hydrocarbon/genetics , Receptors, Aryl Hydrocarbon/metabolism , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Repressor Proteins/chemistry , Repressor Proteins/genetics , Repressor Proteins/metabolismABSTRACT
MAIN CONCLUSION: A basic helix-loop-helix (bHLH) transcription factor gene from Antirrhinum, AmDEL , increases flavonoids accumulation and enhances salt and drought tolerance via up-regulating flavonoid biosynthesis, proline biosynthesis and ROS scavenging genes in transgenic Arabidopsis. In plants, transcriptional regulation is the most important tools for increasing flavonoid biosynthesis. The AmDEL gene, as a basic helix-loop-helix transcription factor gene from Antirrhinum, has been shown to increase flavonoids accumulation in tomato. However, its role in tolerance to abiotic stresses has not yet been investigated. In this study, the codon-optimized AmDEL gene was chemically synthesized. Subcellular localization analysis in onion epidermal cells indicated that AmDEL protein was localized to the nucleus. Expression analysis in yeast showed that the full length of AmDEL exhibited transcriptional activation. Overexpression of AmDEL significantly increased flavonoids accumulation and enhanced salt and drought tolerance in transgenic Arabidopsis plants. Real-time quantitative PCR analysis showed that overexpression of AmDEL resulted in the up-regulation of genes involved in flavonoid biosynthesis, proline biosynthesis and ROS scavenging under salt and drought stresses. Meanwhile, Western blot and enzymatic analyses showed that the activities of phenylalanine ammonia lyase, chalcone isomerase, dihydroflavonol reductase, pyrroline-5-carboxylate synthase, superoxide dismutase and peroxidase were also increased. Further components analyses indicated that the significant increase of proline and relative water content and the significant reduction of H2O2 and malonaldehyde content were observed under salt and drought stresses. In addition, the rates of electrolyte leakage and water loss were reduced in transgenic plants. These findings imply functions of AmDEL in accumulation of flavonoids and tolerance to salt and drought stresses. The AmDEL gene has the potential to be used to increase the content of valuable flavonoids and improve tolerance to abiotic stresses in plants.
Subject(s)
Antirrhinum/genetics , Arabidopsis/genetics , Basic Helix-Loop-Helix Transcription Factors/genetics , Droughts , Flavonoids/metabolism , Plant Proteins/genetics , Salt Tolerance/genetics , Acyltransferases/genetics , Acyltransferases/metabolism , Adaptation, Physiological/genetics , Amino Acid Sequence , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Base Sequence , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/metabolism , Blotting, Western , Cell Nucleus/metabolism , Gene Expression Regulation, Plant , Microscopy, Confocal , Oxidoreductases/genetics , Oxidoreductases/metabolism , Peroxidase/genetics , Peroxidase/metabolism , Phylogeny , Plant Proteins/metabolism , Plants, Genetically Modified , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Superoxide Dismutase/genetics , Superoxide Dismutase/metabolismABSTRACT
Xenopus laevis is a long established model organism for developmental, behavioral and neurological studies. Herein, an updated genome-wide survey was conducted using the ongoing genome project of Xenopus laevis and 106 non-redundant Basic Helix-Loop-Helix (bHLH) genes were identified in the Xenopus laevis genome databases. Gene Ontology (GO) enrichment statistics showed 51 significant GO annotations of biological processes and molecular functions and 5 significant KEGG pathways and a number of Xenopus laevis bHLH genes play significant role in specific development or special physiology processes like the development processes of muscle and eye and other organs. Furthermore, each sub-group of the bHLH family has its special gene functions except for the common GO term categories. Molecular phylogenetic analyses revealed that among these identified bHLH proteins, 105 sequences could classified into 39 families with 46, 25, 10, 5, 16 and 3 members in the corresponding high-order groups A, B, C, D, E and F, respectively with an addition bHLH member categorized as an orphan. The present study provides much useful information for further researches on Xenopus laevis.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/classification , Computational Biology , Databases, Genetic , Xenopus Proteins/classification , Xenopus laevis/classification , Amino Acid Sequence , Animals , Basic Helix-Loop-Helix Transcription Factors/genetics , Basic Helix-Loop-Helix Transcription Factors/metabolism , Evolution, Molecular , Gene Expression Regulation , Genome-Wide Association Study , Molecular Sequence Data , Phylogeny , Xenopus Proteins/genetics , Xenopus Proteins/metabolism , Xenopus laevis/genetics , Xenopus laevis/metabolismABSTRACT
Meiosis marks the transition from the sporophyte to the gametophyte generation in the life cycle of flowering plants, and creates genetic variations through homologous recombination. In most flowering plants, meiosis is highly synchronized within each anther, which is significant for efficient fertilization. To date, little is known about the molecular mechanisms of entry into meiosis and exit from it, and only a few genes in Arabidopsis have been characterized with a role in regulating meiotic progression. In this study, we report the functional characterization of a plant-specific basic helix-loop-helix (bHLH) protein, FEHLSTART (FST), a defect in which leads to premature meiotic entry and asynchronous meiosis, and results in decreased seed yield. Investigation of the time course of meiosis showed that the onset of leptotene, the first stage of prophase I, frequently occurred earlier in fst-1 than in the wild type. Asynchronous meiosis followed, which could manifest in the disruption of regular spindle structures and symmetric cell divisions in fst-1 mutants during the meiosis I/II transition. In accordance with frequently accelerated meiotic entry, whole-transcriptome analysis of fst-1 anthers undergoing meiosis revealed that 19 circadian rhythm genes were affected and 47 pollen-related genes were prematurely expressed at a higher level. Taken together, we propose that FST is required for normal meiotic entry and the establishment of meiotic synchrony.
Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Basic Helix-Loop-Helix Transcription Factors/genetics , Gene Expression Profiling/methods , Meiosis/genetics , Amino Acid Sequence , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/classification , Arabidopsis Proteins/metabolism , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/metabolism , Flowers/genetics , Flowers/growth & development , Flowers/metabolism , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Microscopy, Fluorescence , Molecular Sequence Data , Mutation , Phylogeny , Plants, Genetically Modified , Pollen/genetics , Pollen/growth & development , Pollen/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino AcidABSTRACT
Salvia miltiorrhiza Bunge (Labiatae) is an emerging model plant for traditional medicine, and tanshinones are among the pharmacologically active constituents of this plant. Although extensive chemical and pharmaceutical studies of these compounds have been performed, studies on the basic helix-loop-helix (bHLH) transcription factors that regulate tanshinone biosynthesis are limited. In our study, 127 bHLH transcription factor genes were identified in the genome of S. miltiorrhiza, and phylogenetic analysis indicated that these SmbHLHs could be classified into 25 subfamilies. A total of 19 sequencing libraries were constructed for expression pattern analyses using RNA-Seq. Based on gene-specific expression patterns and up-regulated expression patterns in response to MeJA treatment, 7 bHLH genes were revealed as potentially involved in the regulation of tanshinone biosynthesis. Among them, the gene expression of SmbHLH37, SmbHLH74 and SmbHLH92 perfectly matches the accumulation pattern of tanshinone biosynthesis in S. miltiorrhiza. Our results provide a foundation for understanding the molecular basis and regulatory mechanisms of bHLH transcription factors in S. miltiorrhiza.
Subject(s)
Abietanes/biosynthesis , Basic Helix-Loop-Helix Transcription Factors/genetics , Genes, Plant , Plant Proteins/genetics , Salvia miltiorrhiza/genetics , Acetates/pharmacology , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/metabolism , Cyclopentanes/pharmacology , Oxylipins/pharmacology , Phylogeny , Plant Proteins/classification , Plant Proteins/metabolism , Promoter Regions, Genetic , RNA, Plant/chemistry , RNA, Plant/isolation & purification , RNA, Plant/metabolism , Real-Time Polymerase Chain Reaction , Salvia miltiorrhiza/metabolism , Sequence Analysis, RNA , Transcriptome/drug effectsABSTRACT
Brassinosteroids (BRs) play important roles in many aspects of plant growth and development, including regulation of vascular cambium activities and cell elongation. BR-induced BEE3 (brassinosteroid enhanced expression 3) is required for a proper BR response. Here, we identified a poplar (Populus alba × Populus glandulosa) BEE3-like gene, PagBEE3L, encoding a putative basic helix-loop-helix (bHLH)-type transcription factor. Expression of PagBEE3L was induced by brassinolide (BL). Transcripts of PagBEE3L were mainly detected in stems, with the internode having a low level of transcription and the node having a relatively higher level. The function of the PagBEE3L gene was investigated through phenotypic analyses with PagBEE3L-overexpressing (ox) transgenic lines. This work particularly focused on a potential role of PagBEE3L in stem growth and development of polar. The PagBEE3L-ox poplar showed thicker and longer stems than wild-type plants. The xylem cells from the stems of PagBEE3L-ox plants revealed remarkably enhanced proliferation, resulting in an earlier thickening growth than wild-type plants. Therefore, this work suggests that xylem development of poplar is accelerated in PagBEE3L-ox plants and PagBEE3L plays a role in stem growth by increasing the proliferation of xylem cells to promote the initial thickening growth of poplar stems.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Biomass , Cell Proliferation/genetics , Plant Proteins/genetics , Populus/genetics , Xylem/genetics , Amino Acid Sequence , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/metabolism , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Hybridization, Genetic , Microscopy, Confocal , Molecular Sequence Data , Phylogeny , Plant Proteins/metabolism , Plant Stems/genetics , Plant Stems/growth & development , Plant Stems/metabolism , Plants, Genetically Modified , Populus/growth & development , Populus/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Xylem/cytology , Xylem/metabolismABSTRACT
The basic helix-loop-helix (bHLH) transcription factors are one of the largest families of gene regulatory proteins and play crucial roles in genetic, developmental and physiological processes in eukaryotes. Here, we conducted a survey of the Sus scrofa genome and identified 109 putative bHLH transcription factor members belonging to super-groups A, B, C, D, E, and F, respectively, while four members were orphan genes. We identified 6 most significantly enriched KEGG pathways and 116 most significant GO annotation categories. Further comprehensive surveys in human genome and other 12 medical databases identified 72 significantly enriched biological pathways with these 113 pig bHLH transcription factors. From the functional protein association network analysis 93 hub proteins were identified and 55 hub proteins created a tight network or a functional module within their protein families. Especially, there were 20 hub proteins found highly connected in the functional interaction network. The present study deepens our understanding and provided insights into the evolution and functional aspects of animal bHLH proteins and should serve as a solid foundation for further for analyses of specific bHLH transcription factors in the pig and other mammals.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Genome/genetics , Multigene Family , Sus scrofa/genetics , Amino Acid Sequence , Animals , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/metabolism , Binding Sites/genetics , Evolution, Molecular , Gene Ontology , Humans , Molecular Sequence Data , Phylogeny , Protein Binding , Protein Interaction Maps , Sequence Homology, Amino Acid , Signal Transduction/genetics , Sus scrofa/metabolismABSTRACT
Atonal genes are basic helix-loop-helix transcription factors that were first identified as regulating the formation of mechanoreceptors and photoreceptors in Drosophila. Isolation of vertebrate homologs of atonal genes has shown these transcription factors to play diverse roles in the development of neurons and their progenitors, gut epithelial cells, and mechanosensory cells in the inner ear and skin. In this article, we review the molecular function and regulation of atonal genes and their targets, with particular emphasis on the function of Atoh1 in the development, survival, and function of hair cells of the inner ear. We discuss cell-extrinsic signals that induce Atoh1 expression and the transcriptional networks that regulate its expression during development. Finally, we discuss recent work showing how identification of Atoh1 target genes in the cerebellum, spinal cord, and gut can be used to propose candidate Atoh1 targets in tissues such as the inner ear where cell numbers and biochemical material are limiting.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/physiology , Drosophila Proteins/physiology , Gene Expression Regulation, Developmental/physiology , Mechanoreceptors/physiology , Mechanotransduction, Cellular/physiology , Nerve Tissue Proteins/physiology , Amino Acid Sequence , Animal Structures/physiology , Animals , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/genetics , Compound Eye, Arthropod/cytology , Compound Eye, Arthropod/embryology , Drosophila Proteins/genetics , Drosophila melanogaster/embryology , Drosophila melanogaster/genetics , Drosophila melanogaster/physiology , Evolution, Molecular , Hair Cells, Auditory/physiology , Humans , Intestines/cytology , Intestines/embryology , Intestines/growth & development , Invertebrates/physiology , Merkel Cells/metabolism , Molecular Sequence Data , Nerve Tissue Proteins/genetics , Neurogenesis/physiology , Photoreceptor Cells, Invertebrate/physiology , Phylogeny , Sense Organs/physiology , Sequence Alignment , Sequence Homology, Amino Acid , Signal Transduction/physiology , Transcription, Genetic , Vertebrates/physiologyABSTRACT
Transcription factor is one of the key factors in the regulation of gene expression at the transcriptional level. It plays an important role in plant growth, active components biosynthesis and response to environmental change. This paper summarized the structure and classification of bHLH transcription factors and elaborated the research progress of bHLH transcription factors which regulate the active components in plants, such as flavonoids, alkaloids, and terpenoids. In addition, the possibility of increasing the concentration of active substances by bHLH in medicinal plants was assessed. The paper emphasized great significance of model plants and multidisciplinary research fields including modern genomics, transcriptomics, metabolomics and bioinformatics, providing the contribution to improve the discovery and function characterization of bHLH transcription factors. Accelerating the research in the mechanism of bHLH transcription factors on the regulation of active components biosynthesis will promote the development of breeding and variety improvement of Chinese medicinal materials, also ease the pressure of resources exhaustion of traditional Chinese medicine home and abroad.
Subject(s)
Alkaloids/biosynthesis , Basic Helix-Loop-Helix Transcription Factors/metabolism , Flavonoids/biosynthesis , Plants, Medicinal/metabolism , Basic Helix-Loop-Helix Transcription Factors/chemistry , Basic Helix-Loop-Helix Transcription Factors/classification , Basic Helix-Loop-Helix Transcription Factors/genetics , Plants, Medicinal/genetics , Terpenes/metabolismABSTRACT
Plant basic helix-loop-helix (bHLH) transcription factors (TFs) are involved in a variety of physiological processes including the regulation of plant responses to various abiotic stresses. However, few drought-responsive bHLH family members in Populus have been reported. In this study, a novel bHLH gene (PebHLH35) was cloned from Populus euphratica. Expression analysis in P. euphratica revealed that PebHLH35 was induced by drought and abscisic acid. Subcellular localization studies using a PebHLH35-GFP fusion showed that the protein was localized to the nucleus. Ectopic overexpression of PebHLH35 in Arabidopsis resulted in a longer primary root, more leaves, and a greater leaf area under well-watered conditions compared with vector control plants. Notably, PebHLH35 overexpression lines showed enhanced tolerance to water-deficit stress. This finding was supported by anatomical and physiological analyses, which revealed a reduced stomatal density, stomatal aperture, transpiration rate, and water loss, and a higher chlorophyll content and photosynthetic rate. Our results suggest that PebHLH35 functions as a positive regulator of drought stress responses by regulating stomatal density, stomatal aperture, photosynthesis and growth.