scholarly journals The apple 14-3-3 protein MdGRF11 interacts with the BTB protein MdBT2 to regulate nitrate deficiency-induced anthocyanin accumulation

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Yi-Ran Ren ◽  
Qiang Zhao ◽  
Yu-Ying Yang ◽  
Tian-En Zhang ◽  
Xiao-Fei Wang ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
Anthocyanin Biosynthesis ◽  
Critical Role ◽  
26S Proteasome ◽  
Bimolecular Fluorescence Complementation ◽  
Anthocyanin Accumulation ◽  
The Stability ◽  
Dependent Pathway ◽  
Two Hybrid

AbstractNitrogen is an important factor that affects plant anthocyanin accumulation. In apple, the nitrate-responsive BTB/TAZ protein MdBT2 negatively regulates anthocyanin biosynthesis. In this study, we found that MdBT2 undergoes posttranslational modifications in response to nitrate deficiency. Yeast two-hybrid, protein pull-down, and bimolecular fluorescence complementation (BiFC) assays showed that MdBT2 interacts with MdGRF11, a 14-3-3 protein; 14-3-3 proteins compose a family of highly conserved phosphopeptide-binding proteins involved in multiple physiological and biological processes. The interaction of MdGRF11 negatively regulated the stability of the MdBT2 protein via a 26S proteasome-dependent pathway, which increased the abundance of MdMYB1 proteins to activate the expression of anthocyanin biosynthesis-related genes. Taken together, the results demonstrate the critical role of 14-3-3 proteins in the regulation of nitrate deficiency-induced anthocyanin accumulation. Our results provide a novel avenue to elucidate the mechanism underlying the induction of anthocyanin biosynthesis in response to nitrate deficiency.

scholarly journals MdBBX21, a B-Box Protein, Positively Regulates Light-Induced Anthocyanin Accumulation in Apple Peel

2021 ◽  
Vol 12 ◽  
Author(s):  
Bo Zhang ◽  
Zhen-Zhen Zhu ◽  
Dong Qu ◽  
Bo-Chen Wang ◽  
Ni-Ni Hao ◽  
...  
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Apple Fruit ◽  
Bimolecular Fluorescence Complementation ◽  
Anthocyanin Accumulation ◽  
Rna Seq ◽  
Yeast Two Hybrid ◽  
Fruit Peel ◽  
Apple Peel ◽  
Its Gene ◽  
Two Hybrid

The red coloration of apple (Malus × domestica Borkh.) is due to the accumulation of anthocyanins in the fruit peel. Light is essential for anthocyanin biosynthesis in apple. In this study, we performed a transcriptome sequencing (RNA-seq) analysis of apple fruit exposed to light after unbagging. The identified differentially expressed genes included MdBBX21, which is homologous to Arabidopsis BBX21, suggesting it may be involved in light-induced anthocyanin biosynthesis. Additionally, MdBBX21 was localized in the nucleus and its gene was expressed earlier than MdMYB1 in apple peel treated with light. Overexpressing MdBBX21 in Arabidopsis and apple calli under light increased anthocyanin accumulation. Dual-luciferase and yeast one-hybrid assays confirmed that MdBBX21 binds to the MdHY5, MdBBX20, and MdBBX22-1/2 promoters and induces expression. At the same time, MdHY5 can also activate the expression of MdBBX21. Furthermore, bimolecular fluorescence complementation and yeast two-hybrid assays demonstrated that MdBBX21 can interact with MdHY5. This interaction can significantly enhance MdMYB1 promoter activity. These findings clarify the molecular mechanism by which MdBBX21 positively regulates light-induced anthocyanin accumulation in apple.

scholarly journals SlBBX20 interacts with the COP9 signalosome subunit SlCSN5-2 to regulate anthocyanin biosynthesis by activating SlDFR expression in tomato

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Dan Luo ◽  
Cheng Xiong ◽  
Aihua Lin ◽  
Chunli Zhang ◽  
Wenhui Sun ◽  
...  
Keyword(s):  
Growth Regulation ◽  
Anthocyanin Biosynthesis ◽  
Plant Stress ◽  
Underlying Mechanism ◽  
Bimolecular Fluorescence Complementation ◽  
Cop9 Signalosome ◽  
Anthocyanin Accumulation ◽  
Related Transcription Factor ◽  
Hybrid Screening

AbstractAnthocyanins play vital roles in plant stress tolerance and growth regulation. Previously, we reported that the photomorphogenesis-related transcription factor SlBBX20 regulates anthocyanin accumulation in tomato. However, the underlying mechanism remains unclear. Here, we showed that SlBBX20 promotes anthocyanin biosynthesis by binding the promoter of the anthocyanin biosynthesis gene SlDFR, suggesting that SlBBX20 directly activates anthocyanin biosynthesis genes. Furthermore, we found by yeast two-hybrid screening that SlBBX20 interacts with the COP9 signalosome subunit SlCSN5-2, and the interaction was confirmed by bimolecular fluorescence complementation and coimmunoprecipitation assays. SlCSN5 gene silencing led to anthocyanin hyperaccumulation in the transgenic tomato calli and shoots, and SlCSN5-2 overexpression decreased anthocyanin accumulation, suggesting thSlCSN5-2 enhanced the ubiquitination of SlBBX20 and promoted the degradation of SlBBX20 in vivo. Consistently, silencing the SlCSN5-2 homolog in tobacco significantly increased the accumulation of the SlBBX20 protein. Since SlBBX20 is a vital regulator of photomorphogenesis, the SlBBX20-SlCSN5-2 module may represent a novel regulatory pathway in light-induced anthocyanin biosynthesis.

scholarly journals The role of Clt1-regulated-Xylan metabolism in the melanin and toxin formation for the pathogenicity of Curvularia lunata in Maize

2021 ◽  
Author(s):  
Jinxin Gao ◽  
Jie Chen
Keyword(s):  
Toxin Production ◽  
Bimolecular Fluorescence Complementation ◽  
Curvularia Lunata ◽  
Yeast Two Hybrid ◽  
Acetyl Xylan Esterase ◽  
Btb Domain ◽  
Intermediate Metabolites ◽  
Regulative Mechanism ◽  
Two Hybrid

We previously reported that the BTB domain-containing protein Clt1 regulates melanin and toxin synthesis, conidiation, and pathogenicity in Curvularia lunata, but the interacting proteins and regulative mechanism of Clt1 are unclear. In this research, we identified two proteins, which respectively correspond to xylanase (Clxyn24) and acetyl xylan esterase (Claxe43) from C. lunata were regulated by Clt1. Yeast two-hybrid (Y2H), and bimolecular fluorescence complementation assays were conducted to verify the interaction of Clt1 with full-length Clxyn24 and Claxe43. Furthermore, the Y2H assay revealed that Clt1 physically interacted with Clxyn24 and Claxe43 through its BTB domain to degrade xylan which was used as a carbon source for C. lunata growth. The utilization of xylan provides acetyl-CoA for the synthesis of melanin and toxin, as well as energy and other intermediate metabolites for conidiation. Furthermore, transcriptome analysis revealed that PKS18 and its 13 flanking genes are found clustered in a region spanning 57.89 kb on scaffold 9 of the C. lunata CX-3 genome were down-regulated in toxin production deficient mutant T806, and this cluster is possibly responsible for toxin biosynthesis of C. lunata.

Role of miRNAs In Cancer Diagnostics And Therapy: A Recent Update

2021 ◽  
Vol 27 ◽  
Author(s):  
Adil A. Sayyed ◽  
Piyush Gondaliya ◽  
Palak Bhat ◽  
Mukund Mali ◽  
Neha Arya ◽  
...  
Keyword(s):  
Targeted Delivery ◽  
Tumor Invasion ◽  
Cancer Metastasis ◽  
Critical Role ◽  
Delivery Systems ◽  
Cancer Diagnostics ◽  
Disease Monitoring ◽  
The Stability ◽  
Diagnostics And Therapy

: The discovery of miRNAs has been one of the revolutionary developments and has led to the advent of new diagnostic and therapeutic opportunities for the management of cancer. In this regard, miRNA dysregulation has been shown to play a critical role in various stages of tumorigenesis, including tumor invasion, metastasis as well as angiogenesis. Therefore, miRNA profiling can provide accurate fingerprints for the development of diagnostic and therapeutic platforms. This review discusses the recent discoveries of miRNA-based tools for early detection of cancer as well as disease monitoring in cancers that are common, like breast, lung, hepatic, colorectal, oral and brain cancer. Based on the involvement of miRNA in different cancers as oncogenic miRNA or tumor suppressor miRNA, the treatment with miRNA inhibitors or mimics is recommended. However, the stability and targeted delivery of miRNA remain the major limitations of miRNA delivery. In relation to this, several nanoparticle-based delivery systems have been reported which have effectively delivered the miRNA mimics or inhibitors and showed the potential for transforming these advanced delivery systems from bench to bedside in the treatment of cancer metastasis and chemoresistance. Based on this, we attempted to uncover recently reported advanced nanotherapeutic approaches to deliver the miRNAs in the management of different cancers.

scholarly journals Viperin catalyzes methionine oxidation to promote protein expression and function of helicases

2019 ◽  
Vol 5 (8) ◽  
pp. eaax1031 ◽  
Author(s):  
Lei Bai ◽  
Jiazhen Dong ◽  
Zhenqiu Liu ◽  
Youliang Rao ◽  
Pinghui Feng ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
Methionine Oxidation ◽  
Biological Processes ◽  
Loss Of Function ◽  
Rna Helicases ◽  
Dna And Rna ◽  
Biochemical Studies ◽  
The Stability ◽  
And Function

Helicases play pivotal roles in fundamental biological processes, and posttranslational modifications regulate the localization, function, and stability of helicases. Here, we report that methionine oxidation of representative helicases, including DNA and RNA helicases of viral (ORF44 of KSHV) and cellular (MCM7 and RIG-I) origin, promotes their expression and functions. Cellular viperin, a major antiviral interferon-stimulated gene whose functions beyond host defense remain largely unknown, catalyzes the methionine oxidation of these helicases. Moreover, biochemical studies entailing loss-of-function mutations of helicases and a pharmacological inhibitor interfering with lipid metabolism and, hence, decreasing viperin activity indicate that methionine oxidation potently increases the stability and enzyme activity of these helicases that are critical for DNA replication and immune activation. Our work uncovers a pivotal role of viperin in catalyzing the methionine oxidation of helicases that are implicated in diverse fundamental biological processes.

scholarly journals Excessive Neutrophil Extracellular Trap Formation Aggravates Acute Myocardial Infarction Injury in Apolipoprotein E Deficiency Mice via the ROS-Dependent Pathway

2019 ◽  
Vol 2019 ◽  
pp. 1-15 ◽  
Author(s):  
Zheliang Zhou ◽  
Shuning Zhang ◽  
Suling Ding ◽  
Mieradilijiang Abudupataer ◽  
Zhiwei Zhang ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Apolipoprotein E ◽  
Myocardial Injury ◽  
Early Stage ◽  
Critical Role ◽  
Neutrophil Extracellular Trap ◽  
Coronary Artery Ligation ◽  
Infarct Zone ◽  
Dependent Pathway

Genetically human apolipoprotein E (APOE) ε32 is associated with a decreased risk of ischemic heart disease. ApoE deficiency in mice impairs infarct healing after myocardial infarction (MI). After the ischemic injury, a large number of neutrophils are firstly recruited into the infarct zone and then degrade dead material and promote reparative phase transformation. The role of ApoE in inflammation response in the early stage of MI remains largely unclear. In this study, we investigated the effect of ApoE deficiency on neutrophils’ function and myocardial injury after myocardial infarction. By left coronary artery ligation in ApoE-/- and wild-type (WT) mice, we observed increased infarct size and neutrophil infiltration in ApoE-/- mice. Within the infarct zone, more neutrophil extracellular traps (NETs) were observed in ApoE-/- mice, while increased ex vivo NET formation was detected in ApoE-/- mouse-derived neutrophils through the NADPH oxidase-ROS-dependent pathway. Suppressing overproduced NETs reduced myocardial injury in ApoE-/- mice after ligation. In general, our findings reveal a critical role of apolipoprotein E in regulating Ly6G+ neutrophil activation and NET formation, resulting in limiting myocardial injury after myocardial infarction. In such a process, apolipoprotein E regulates NET formation via the ROS-MAPK-MSK1 pathway.

scholarly journals Mutational Analysis of VIM-2 Reveals an Essential Determinant for Metallo-β-Lactamase Stability and Folding

2010 ◽  
Vol 54 (8) ◽  
pp. 3197-3204 ◽  
Author(s):  
Luisa Borgianni ◽  
Julie Vandenameele ◽  
André Matagne ◽  
Luca Bini ◽  
Robert A. Bonomo ◽  
...  
Keyword(s):  
Mutational Analysis ◽  
Critical Role ◽  
Kinetic Studies ◽  
Amino Acid Position ◽  
Structural Knowledge ◽  
Saturation Mutagenesis ◽  
Biochemical Analyses ◽  
The Stability ◽  
Substrate Hydrolysis

ABSTRACT Metallo-β-lactamase (MBL)-producing bacteria are emerging worldwide and represent a formidable threat to the efficacy of relevant β-lactams, including carbapenems, expanded-spectrum cephalosporins, and β-lactamase inactivator/β-lactam combinations. VIM-2 is currently the most widespread MBL and represents a primary target for MBL inhibitor research, the clinical need for which is expected to further increase in the future. Using a saturation mutagenesis approach, we probed the importance of four residues (Phe-61, Ala-64, Tyr-67, and Trp-87) located close to the VIM-2 active site and putatively relevant to the enzyme activity based on structural knowledge of the enzyme and on structure-activity relationships of the subclass B1 MBLs. The ampicillin MIC values shown by the various mutants were affected very differently depending on the randomized amino acid position. Position 64 appeared to be rather tolerant to substitution, and kinetic studies showed that the A64W mutation did not significantly affect substrate hydrolysis or binding, representing an important difference from IMP-type enzymes. Phe-61 and Tyr-67 could be replaced with several amino acids without the ampicillin MIC being significantly affected, but in contrast, Trp-87 was found to be critical for ampicillin resistance. Further kinetic and biochemical analyses of W87A and W87F variants showed that this residue is apparently important for the structure and proper folding of the enzyme but, surprisingly, not for its catalytic activity. These data support the critical role of residue 87 in the stability and folding of VIM-2 and might have strong implications for MBL inhibitor design, as this residue would represent an ideal target for interaction with small molecules.

scholarly journals The Basic Helix-Loop-Helix Transcription Factor SmbHLH1 Represses Anthocyanin Biosynthesis in Eggplant

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhaofei Duan ◽  
Shiyu Tian ◽  
Guobin Yang ◽  
Min Wei ◽  
Jing Li ◽  
...  
Keyword(s):  
Plant Species ◽  
Anthocyanin Biosynthesis ◽  
Solanum Melongena ◽  
Amino Acid Sequences ◽  
Negative Regulator ◽  
Bimolecular Fluorescence Complementation ◽  
Luciferase Assay ◽  
Anthocyanin Accumulation ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix

Many basic helix-loop-helix transcription factors (TFs) have been reported to promote anthocyanin biosynthesis in numerous plant species, but little is known about bHLH TFs that inhibit anthocyanin accumulation. In this study, SmbHLH1 from Solanum melongena was identified as a negative regulator of anthocyanin biosynthesis. However, SmbHLH1 showed high identity with SmTT8, which acts as a SmMYB113-dependent positive regulator of anthocyanin-biosynthesis in plants. Overexpression of SmbHLH1 in eggplant caused a dramatic decrease in anthocyanin accumulation. Only the amino acid sequences at the N and C termini of SmbHLH1 differed from the SmTT8 sequence. Expression analysis revealed that the expression pattern of SmbHLH1 was opposite to that of anthocyanin accumulation. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays showed that SmbHLH1 could not interact with SmMYB113. Dual-luciferase assay demonstrated that SmbHLH1 directly repressed the expression of SmDFR and SmANS. Our results demonstrate that the biological function of bHLHs in anthocyanin biosynthesis may have evolved and provide new insight into the molecular functions of orthologous genes from different plant species.

scholarly journals The RNA Directed DNA Methylation (RdDM) Pathway Regulates Anthocyanin Biosynthesis in Crabapple (Malus cv. spp.) Leaves by Methylating the McCOP1 Promoter

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2466
Author(s):  
Yifan Xing ◽  
Ziyi Xie ◽  
Weilei Sun ◽  
Yuying Sun ◽  
Zhenyun Han ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Methylation Level ◽  
Bisulfite Sequencing ◽  
Anthocyanin Biosynthesis ◽  
Apple Fruit ◽  
Bimolecular Fluorescence Complementation ◽  
Rna Expression ◽  
Anthocyanin Accumulation ◽  
Transcript Levels ◽  
Direct Target

The synthesis of anthocyanin pigments in plants is known to be regulated by multiple mechanisms, including epigenetic regulation; however, the contribution of the RNA-directed DNA methylation (RdDM) pathway is not well understood. Here, we used bisulfite sequencing and Real Time (RT)-quantitative (q) PCR to analyze the methylation level of the promoter of constitutively photomorphogenic 1 (McCOP1) from Malus cv. spp, a gene involved in regulating anthocyanin biosynthesis. The CHH methylation level of the McCOP1 promoter was negatively correlated with McCOP1 RNA expression, and inhibiting DNA methylation caused decreased methylation of the McCOP1 promoter and asymmetric cytosine CHH methylation. We observed that the McCOP1 promoter was a direct target of the RdDM pathway argonaute RISC component 4 (McAGO4) protein, which bound to a McCOP1 promoter GGTTCGG site. Bimolecular fluorescence complementation (BIFC) analysis showed that RNA-directed DNA methylation (McRDM1) interacted with McAGO4 and another RdDM protein, domains rearranged methyltransferase 2 (McDRM2), to regulate the CHH methylation of the McCOP1 promoter. Detection of CHH methylation and COP1 gene expression in the Arabidopsis thalianaatago4, atdrm2 and atrdm1 mutants showed that RDM1 is the effector of the RdDM pathway. This was confirmed by silencing McRDM1 in crabapple leaves or apple fruit, which resulted in a decrease in McCOP1 CHH methylation and an increase in McCOP1 transcript levels, as well as in anthocyanin accumulation. In conclusion, these results show that the RdDM pathway is involved in regulating anthocyanin accumulation through CHH methylation of the McCOP1 promoter.

scholarly journals Fundamental Role Of The H2A.Z C-Terminal Tail In The Formation Of Constitutive Heterochromatin

2021 ◽  
Author(s):  
László Imre ◽  
Péter Nánási ◽  
Rosevalentine Bosire ◽  
Ágota Csóti ◽  
Kata Nóra Enyedi ◽  
...  
Keyword(s):  
Posttranslational Modifications ◽  
Constitutive Heterochromatin ◽  
Large Fraction ◽  
Gene Expression Pattern ◽  
Histone Variant ◽  
Unusual Behavior ◽  
Isoform Specificity ◽  
The Stability

ABSTRACTNucleosome stability, a crucial determinant of gene regulation, was measured in a robust in situ assay to assess the molecular determinants of the stability of H2A.Z-containig nucleosomes. Surprisingly, a large fraction of H2A.Z detected by three different antibodies was released from the nucleosomes by salt together with H3, and was associated with H3K9me3 but not with H3K27me3 marked nucleosomes. This unusual behavior relied on the presence of the unstructured C-terminal chain of the histone variant, rather than on isoform specificity, posttranslational modifications or binding of the reader protein PWWPA2, as determined using cell lines expressing only particular forms of the variant. In the absence of this tail, or upon addition of an excess of the tail peptide to the nuclei of control cells, the canonical H2A-like stability features were readily restored and most of the H2A.Z-containing nucleosomes left the periphery and ended up in scattered foci in the nuclei. Concomitantly, the H3K9me3-marked constitutive heterochromatin was also dispersed, what was accompanied by increased overall nuclease sensitivity and significantly enhanced binding of intercalating dyes to the DNA. The DT40 cells expressing the tailless H2A.Z showed marked differences in their gene expression pattern and were distinguished by compromised DNA damage response. Thus, interactions involving a short H2A.Z peptide chain simultaneously determine the stability and accessibility features of chromatin involving the nucleosomes containing this histone variant and the localization of these large chromatin regions in the nucleus. Our data suggest that H2A.Z can function in both heterochromatic and in euchromatic scenarios depending on the molecular interactions involving its C-terminal unstructured tail, shedding light on the enigmatic double-faced character of this histone variant.

Sign in / Sign up

Export Citation Format

Share Document