1.Phylogenetic and expression analysis of SEPALLATA-like gene in Brassica oleracea L. var. acephala.
Yuanping XIANG ; Yuntong HUANG ; Hongjun HE ; Qijiang XU
Chinese Journal of Biotechnology 2020;36(11):2398-2412
The E class MADS-box genes SEPALLATA (SEP)-like play critical roles in angiosperm reproductive growth, especially in floral organ differentiation. To analyze the sequence characteristics and spatio-temporal expression patterns of E-function MADS-box SEP-like genes during kale (Brassica oleracea L. var. acephala) flower development, BroaSEP1/2/3 (GenBank No. KC967957, KC967958, KC967960) homologues, three kale SEP MADS-box gene, were isolated from the kale variety 'Fourteen Line' using Rapid amplification of cDNA ends (RACE). Sequence and phylogenetic analysis indicated that these three SEP genes had a high degree of identity with SEP1, SEP2, SEP3 from Brassica oleracea var. oleracea, Brassica rapa, Raphanus sativus and Brassica napus, respectively. Alignment of the predicted amino acid sequences from these genes, along with previously published subfamily members, demonstrated that these genes comprise four regions of the typical MIKC-type MADS-box proteins: the MADS domain, intervening (I) domain and keratin-like (K) domain, and the C-terminal domain SEPⅠ and SEP Ⅱ motif. The longest open reading frame deduced from the cDNA sequences of BroaSEP1, BroaSEP2, and BroaSEP3 appeared to be 801 bp, 759 bp, 753 bp in length, respectively, which encoded proteins of 266, 252, and 250 amino acids respectively. Expression analyses using semi-quantitative RT-PCR and quantitative real-time PCR indicate that BroaSEP1/2/3 are specifically expressed in floral buds of kale during flower development process. The expression levels of the three genes are very different at different developmental stages, also in wild type, mutant flower with increased petals, and mutant flower with decreased petals. These different patterns of gene expression maybe cause the flowers to increase or decrease the petal number.
Brassica/metabolism*
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Flowers/genetics*
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Gene Expression Regulation, Plant
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MADS Domain Proteins/metabolism*
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Phylogeny
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Plant Proteins/metabolism*
2.Identification and expression analysis of MADS-box gene family in Docynia delavayi (Franch.) Schneid.
Xiwei WANG ; Can CHEN ; Dawei WANG
Chinese Journal of Biotechnology 2023;39(7):2897-2913
MADS-box gene family is a significant transcription factor family that plays a crucial role in regulating plant growth, development, signal transduction, and other processes. In order to study the characteristics of MADS-box gene family in Docynia delavayi (Franch.) Schneid. and its expression during different stages of seed germination, this study used seedlings at different stages of germination as materials and screened MADS-box transcription factors from the transcriptome database of D. delavayi using bioinformatics methods based on transcriptome sequencing. The physical and chemical properties, protein conservative motifs, phylogenetic evolution, and expression patterns of the MADS-box transcription factors were analyzed. Quantitative real-time PCR (qRT-PCR) was used to verify the expression of MADS-box gene family members during different stages of seed germination in D. delavayi. The results showed that 81 genes of MADS-box gene family were identified from the transcriptome data of D. delavayi, with the molecular weight distribution ranged of 6 211.34-173 512.77 Da and the theoretical isoelectric point ranged from 5.21 to 10.97. Phylogenetic analysis showed that the 81 genes could be divided into 15 subgroups, among which DdMADS27, DdMADS42, DdMADS45, DdMADS46, DdMADS53, DdMADS61, DdMADS76, DdMADS77 and DdMADS79 might be involved in the regulation of ovule development in D. delavayi. The combination of the transcriptome data and the qRT-PCR analysis results of D. delavayi seeds indicated that DdMADS25 and DdMADS42 might be involved in the regulation of seed development, and that DdMADS37 and DdMADS38 might have negative regulation effects on seed dormancy. Previous studies have reported that the MIKC* subgroup is mainly involved in regulating flower organ development. For the first time, we found that the transcription factors of the MIKC* subgroup exhibited a high expression level at the early stage of seed germination, so we speculated that the MIKC* subgroup played a regulatory role in the process of seed germination. To verify the accuracy of this speculation, we selected DdMADS60 and DdMADS75 from the MIKC* subgroup for qRT-PCR experiments, and the experimental results were consistent with the expression trend of transcriptome sequencing. This study provides a reference for further research on the biological function of D. delavayi MADS-box gene family from the perspective of molecular evolution.
MADS Domain Proteins/metabolism*
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Phylogeny
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Gene Expression Regulation, Plant
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Genes, Plant
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Transcription Factors/genetics*
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Plant Proteins/metabolism*
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Gene Expression Profiling
3.The enzymatic activity of Arabidopsis protein arginine methyltransferase 10 is essential for flowering time regulation.
Lifang NIU ; Falong LU ; Taolan ZHAO ; Chunyan LIU ; Xiaofeng CAO
Protein & Cell 2012;3(6):450-459
Arabidopsis AtPRMT10 is a plant-specific type I protein arginine methyltransferase that can asymmetrically dimethylate arginine 3 of histone H4 with auto-methylation activity. Mutations of AtPRMT10 derepress FLOWERING LOCUS C (FLC) expression resulting in a late-flowering phenotype. Here, to further investigate the biochemical characteristics of AtPRMT10, we analyzed a series of mutated forms of the AtPRMT10 protein. We demonstrate that the conserved "VLD" residues and "double-E loop" are essential for enzymatic activity of AtPRMT10. In addition, we show that Arg54 and Cys259 of AtPRMT10, two residues unreported in animals, are also important for its enzymatic activity. We find that Arg13 of AtPRMT10 is the auto-methylation site. However, substitution of Arg13 to Lys13 does not affect its enzymatic activity. In vivo complementation assays reveal that plants expressing AtPRMT10 with VLD-AAA, E143Q or E152Q mutations retain high levels of FLC expression and fail to rescue the late-flowering phenotype of atprmt10 plants. Taken together, we conclude that the methyltransferase activity of AtPRMT10 is essential for repressing FLC expression and promoting flowering in Arabidopsis.
Arabidopsis
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enzymology
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Arabidopsis Proteins
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biosynthesis
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genetics
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metabolism
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Enzyme Activation
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Flowers
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genetics
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growth & development
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metabolism
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Genetic Loci
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genetics
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MADS Domain Proteins
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biosynthesis
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genetics
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metabolism
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Methyltransferases
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genetics
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metabolism
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Phenotype
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Recombinant Proteins
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genetics
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metabolism
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Time Factors
4.Molecular cloning and spatiotemporal expression of an APETALA1/FRUITFULL-like MADS-box gene from the orchid (Cymbidium faberi).
Yunfang TIAN ; Xiuyun YUAN ; Suhua JIANG ; Bo CUI ; Jinle SU
Chinese Journal of Biotechnology 2013;29(2):203-213
In order to identify genes involved in floral transition and development of the orchid species, a full-length APETALA1/FRUITFULL-like (AP1/FUL-like) MADS box cDNA was cloned from Cymbidium faberi (C. faberi) sepals and designated as C. faberi APETALA1-like (CfAP11], JQ031272.1). The deduced amino acid sequence of CfAP11 shared 84% homology with a member of the AP1/FUL-like group of MADS box genes (AY927238.1, Dendrobium thyrsiflorum FUL-like MADS box protein 3 mRNA). Phylogenetic analysis shows that CfAP11 belonged to the AP1/FUL transcription factor subfamily. Bioinformatics analysis shows that the deduced protein had a MADS domain and a relatively conservative K region. The secondary structure of CfAP11 mainly consisted of alpha helices (58.97%), and the three-dimensional structure of the protein was similar to that of homologues in Roza hybrida, Oryza sativa and Narcissus tazetta. Real-time quantitative PCR (qRT-PCR) results reveal low levels of its mRNA in roots, lower levels in leaves during reproductive period than vegetative period, and higher levels in pedicels at full-blossom stage than at bud stage. These results suggest that CfAP11 is involved in floral induction and floral development. Additionally, we observed higher levels of CfAP11 expression in pedicels and ovaries than in other tissues during full-blossom stage, which suggests that CfAP11 may also be involved in fruit formation in certain mechanism.
Amino Acid Sequence
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Cloning, Molecular
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Flowers
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genetics
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metabolism
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Gene Expression Regulation, Plant
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Genes, Plant
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MADS Domain Proteins
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genetics
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Molecular Sequence Data
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Orchidaceae
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genetics
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metabolism
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Plant Proteins
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genetics
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metabolism
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Spatio-Temporal Analysis
5.Expression of MEF2D on nasopharyngeal carcinoma tissues and its influence of prognostic.
Yongling LI ; Longcheng ZHANG ; Jiang NONG ; Shixia BIAN ; Zhen ZHAO ; Yi REN ; Xinran LIN ; Xiuwu BIAN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2011;25(18):840-847
OBJECTIVE:
To explore the expression of MEF2D in NPC tissues, study the relationship between the expression and prognostic.
METHOD:
Specimens from 101 NPC patients who were follow-up visited 1 to 7 years were analyzed for MEF2D by using immunohistochemistry.
RESULT:
(1) The expression of MEF2D was higher in the higher clinical stage. (2) Density and Grey of MEF2D was negative correlated (|r| = 0.865, P < 0.01). (3) NPC patients' survival rate after therapies was 52.5%, the survival curve of 1th clinical stage was higher than 4th. (4) The survival curves of MEF2D stages were no statistical significance.
CONCLUSION
There's statistical significance of the MEF2D expression in clinical stages, but not in survival curve, which indicated that MEF2D concerned with invasion and metastatic of NPC.
Adult
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Carcinoma
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Carcinoma, Squamous Cell
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metabolism
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pathology
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Female
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Humans
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Lymphatic Metastasis
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MADS Domain Proteins
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metabolism
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MEF2 Transcription Factors
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Male
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Middle Aged
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Myogenic Regulatory Factors
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metabolism
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Nasopharyngeal Carcinoma
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Nasopharyngeal Neoplasms
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metabolism
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pathology
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Neoplasm Staging
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Prognosis
6.Regulation pattern of the FRUITFULL (FUL) gene of Arabidopsis thaliana.
Tingting CHU ; Hua XIE ; Yong XU ; Rongcai MA
Chinese Journal of Biotechnology 2010;26(11):1546-1554
FRUITFULL (FUL) is an MADS box gene that functions early in controlling flowering time, meristem identity and cauline leaf morphology and later in carpel and fruit development in Arabidopsis thaliana. In order to clarify the regulation of FUL expression the upstream regulatory region, -2148 bp - +96 bp and the first intron of the FUL gene were cloned, and vectors with a series of deletion of FUL promoter, and the ones fused with the first intron were constructed. Vectors harboring the fusion of cis-acting elements with the constitutive promoters of TUBULIN and ACTIN were also constructed. Beta-Glucuronidase activity assays of the transgenic Arabidopsis plants showed that two cis-elements were involved in the repression of FUL expression, with one of the two being probably the binding site of the transcriptional factor AP1. And the two CArG boxes played a important role in FUL initiation particularly. Furthermore, the first intron of FUL was shown to participate in the development of carpel and stamen as an enhancer.
Actins
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genetics
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Arabidopsis
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genetics
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metabolism
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Arabidopsis Proteins
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genetics
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Base Sequence
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Enhancer Elements, Genetic
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Flowers
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genetics
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metabolism
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Gene Expression Regulation, Plant
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Introns
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genetics
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MADS Domain Proteins
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genetics
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Molecular Sequence Data
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Promoter Regions, Genetic
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genetics
7.Function of plant homeodomain-finger proteins in vernalization pathway in Arabidopsis and other cruciferous plants.
Gongling HU ; Guoping CHEN ; Zongli HU ; Feng GU ; Yong LI
Chinese Journal of Biotechnology 2010;26(1):1-8
Vernalization makes Arabidopsis and other cruciferous plants flowering earlier. During this process, an important plant homeodomain-finger(PHD-finger) protein named VIN3 is involved. The PHD domain was a conserved zinc-finger domain in eukaryotic organism. It used to take part in the interaction between proteins, especially the modification on histone of nucleosome, such as methylation, acetylation and phosphorylation. In vernaliazation pathway, the proteins translated by VERNALIZATION INSENSITIVE 3(VIN3) and homologous genes could result in methylation on H3K9 and H3K27 and deacetylation on H3K9 and H3K14 on chromatin histone of FLOWERING LOCUS C, a gene that inhibited flowering. The structure state of FLC would be changed from relaxation into compression. Then the transcription activity of FLC could be restrained and it couldn't inhibit flowering any more, so it would induce flowering earlier. This paper reviewed the function of PHD-finger proteins in vernalization pathway in Arabidopsis and other cruciferous plants, and overviewed the vernalization mechanism.
Amino Acid Sequence
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Arabidopsis
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genetics
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metabolism
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Arabidopsis Proteins
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genetics
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metabolism
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physiology
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Brassicaceae
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genetics
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DNA-Binding Proteins
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genetics
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metabolism
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physiology
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Gene Expression Regulation, Plant
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genetics
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physiology
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Histones
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metabolism
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Homeodomain Proteins
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genetics
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metabolism
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physiology
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MADS Domain Proteins
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genetics
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metabolism
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Molecular Sequence Data
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Transcription Factors
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genetics
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metabolism
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physiology
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Zinc Fingers
8.Understanding the genetic and epigenetic architecture in complex network of rice flowering pathways.
Changhui SUN ; Dan CHEN ; Jun FANG ; Pingrong WANG ; Xiaojian DENG ; Chengcai CHU
Protein & Cell 2014;5(12):889-898
Although the molecular basis of flowering time control is well dissected in the long day (LD) plant Arabidopsis, it is still largely unknown in the short day (SD) plant rice. Rice flowering time (heading date) is an important agronomic trait for season adaption and grain yield, which is affected by both genetic and environmental factors. During the last decade, as the nature of florigen was identified, notable progress has been made on exploration how florigen gene expression is genetically controlled. In Arabidopsis expression of certain key flowering integrators such as FLOWERING LOCUS C (FLC) and FLOWERING LOCUS T (FT) are also epigenetically regulated by various chromatin modifications, however, very little is known in rice on this aspect until very recently. This review summarized the advances of both genetic networks and chromatin modifications in rice flowering time control, attempting to give a complete view of the genetic and epigenetic architecture in complex network of rice flowering pathways.
Arabidopsis
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genetics
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growth & development
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metabolism
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Arabidopsis Proteins
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genetics
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metabolism
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Chromatin
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chemistry
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metabolism
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Epigenesis, Genetic
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Florigen
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metabolism
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Flowers
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genetics
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growth & development
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metabolism
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Gene Expression Regulation, Plant
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Gene Regulatory Networks
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MADS Domain Proteins
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genetics
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metabolism
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Oryza
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genetics
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growth & development
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metabolism
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Phenotype
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Time Factors
9.Study of the 482G/A variation in PGC-1alpha gene domain MEF2C as possible mechanism of type 2 diabetes.
Wensheng LU ; Xiaodong YAN ; Qin HUANG ; Yingyu HU ; Mei ZHONG ; Zhong HUANG ; Hui CHEN ; Hua CHENG
Chinese Journal of Medical Genetics 2008;25(6):616-623
OBJECTIVETo investigate the association of the 482G/A polymorphism of the PGC-1alpha gene with type 2 diabetes by family-based study in the Han population in South China, and to analyze the quantitative and qualitative binding force changes between the PGC-1alpha domain mutant and MEF2C, as well as to evaluate the possibility of PGC-1alpha -MEF2C-GLUT4 pathway in the pathogenesis of type 2 diabetes.
METHODSBlood samples were collected from 350 patients with type 2 diabetes and their first-degree relatives. Genomic DNA was extracted and polymorphic PGC-1alpha genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism and direct DNA sequencing. The results were analyzed by family-based transmission disequilibrium test (TDT) and haplotype relative risk (HRR). The protein-protein interaction between PGC-1alpha and MEF2C was detected by means of the site-directed mutagenesis kit and bacteriomatch two-hybrid system kit.
RESULTSIn the family-based study, HRR analyses demonstrated that the 482A allele was more often transmitted to patients than predicted by chance (chi (2)= 7.2170, P= 0.0072, HRR= 1.4496). TDT-extended test(ETDT) analyses also revealed that PGC-1alpha 482A allele was significantly deviated from 0.5 from heterozygous parents to patients than expected (219 trios, P= 0.0310; 350 trios, P= 0.0292). BacterioMatch Two-Hybrid System showed that 482A variation could lead to decreased binding force between PGC-1alpha and MEF2C (62.1+/- 8.97, P< 0.05).
CONCLUSIONThe 482A polymorphism increases the risk of developing type 2 diabetic mellitus in the South China Han population, which might be mediated by the PGC-1alpha -MEF2C-GLUT4 pathway.
Asian Continental Ancestry Group ; genetics ; Diabetes Mellitus, Type 2 ; genetics ; metabolism ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Glucose Transporter Type 4 ; metabolism ; Haplotypes ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Logistic Models ; MADS Domain Proteins ; genetics ; metabolism ; MEF2 Transcription Factors ; Male ; Middle Aged ; Myogenic Regulatory Factors ; genetics ; metabolism ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Polymorphism, Single Nucleotide ; genetics ; Protein Structure, Tertiary ; genetics ; Signal Transduction ; Transcription Factors ; genetics ; metabolism ; Two-Hybrid System Techniques