Horticulturae
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2022
·Vol. 8
·期 11
·1024
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IF 2.923
Tao Xu,Tianchi Chen,Tianye Zhang,Leyi Shen,Zhe Chen,Yue Xu,Yueyan Wu,Jian Yang
With the frequent occurrence of extreme natural disasters, unfavorable growth environment is a common phenomenon in the life cycle of plants. In recent years, universal stress proteins (USPs) have attracted extensive attention in the field of plant science for their expression patterns and functional analysis. However, the characterization of the USP family remains unclear in grape. In this study, we identified 30VvUSPsin the grape genome, which could be divided into three classes according to their encoded protein sequences, and this classification was reflected by the distribution of conserved motifs. Gene duplication analysis demonstrated that segmental duplication was an important pathway in the expansion of theVvUSPfamily. The expression patterns of 12VvUSPswere significantly different between tissues, implying they had different functions in various tissues.Cis-acting element and expression analysis showed that most of the promoter regions ofVvUSPscontained sequences responsive to hormones and stress elements, especially the promoter region of VIT_16s0013g01920. In conclusion, our findings provide comprehensive information for the further investigation of the genetics and protein functions of theUSPgene family in grape.Keywords:grape;universal stress protein;abiotic stress;hormone
研究领域:
植物科学
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基因组学
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分子生物学
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DOI
高影响因子
MICROCHIMICA ACTA
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2023
·Vol. 190
·期 5
·1-9
·
IF 5.7
Li Xinxin,Zhang Meiling,Zhang Hua,Wang Zhenxin,Zhang Huimao
A simple fluorescence resonance energy transfer (FRET) sensing platform (termed as USP), comprised of upconversion nanoparticles (UCNPs) as the energy donor and Cy5 as the energy acceptor, has been synthesized for cathepsin B (CTSB) activity detection in vitro and in vivo. When Cy5-modified peptide substrate (peptide-Cy5) of CTSB is covalently linked on the surface of UCNPs, the FRET between the UCNPs (excitation: 980 nm; emission: 541 nm/655 nm) and Cy5 (excitation: 645 nm) leads to a reduction in the red upconversion luminescence (UCL) signal intensity of UCNPs. Cy5 can be liberated from UCNPs in the presence of CTSB through the cleavage of peptide-Cy5 by CTSB, leading to the recovery of the red UCL signal of UCNPs. Because the green UCL signal of UCNPs remains constant during the CTSB digestion, it can be considered as an internal reference. The findings demonstrate the ability of USP to detect CTSB with the linear detection ranges of 1 to 100 ng·mL −1 in buffer and 2 × 10 3 to 1 × 10 5 cells in 0.2 mL cell lysates. The limits of detection (LODs) are 0.30 ng·mL −1 in buffer and 887 cells in 0.2 mL of cell lysates (S/N = 3). The viability of USP to detect CTSB activity in tumor-bearing mice is has further been investigated using in vivo fluorescent imaging. Graphical abstract
研究领域:
生物技术
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生物化学
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分子生物学
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癌症研究
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纳米技术
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DOI
PubMed
高影响因子
BIOCHEMICAL PHARMACOLOGY
·
2024
·
IF 5.3
Hongzhuan Yin,Di Wu,Qiao Qu,Zhilong Li,Lianrong Zhao
The tumor recurrence and metastasis of colorectal cancer (CRC) are responsible for most of CRC-linked mortalities. It is an urgent need to deeply investigate the pathogenesis of CRC metastasis and look for novel targets for its treatment. The current study aimed to investigate the effects of ubiquitin-specific peptidase 15 (USP-15) on the CRC progression. In vivo, a mouse model of liver metastasis of CRC tumor was established to investigate the role of USP-15. In vitro, the migrated and invasive abilities of CRC cells were assessed by transwell assay. Cell stemness was evaluated by using sphere formation assay. The underlying mechanism was further explored by employing the co-immunoprecipitation, dual luciferase reporter assay, oligonucleotide pull-down assay, and chromatin immunoprecipitation assay. The results showed that USP-15 was upregulated in CRC patients with liver metastasis and high metastatic potential cell lines of CRC. Loss of USP-15 repressed the epithelial-to-mesenchymal transition (EMT), migration, invasion, and stemness properties of CRC cells in vitro. Downregulation of USP-15 reduced the liver metastasis of mice in vivo. USP-15 upregulation obtained the contrary effects. Subsequently, USP-15 deubiquitinated transcription factor AP-4 (TFAP4) and enhanced its protein stability. TFAP4 could transcriptionally activated polycomb group ring finger 1 (PCGF1). The pro-cancer effects of USP-15 were rescue by the knockdown of TFAP4 or PCGF1. In conclusions: USP-15 facilitated the liver metastasis by the enhancement of cell stemness and EMT in CRC, which was at least partly mediated by the deubiquitination of TFAP4 upon the upregulation of PCGF1.
研究领域:
肿瘤学
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分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
Nature Protocols
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2024
·
IF 13.1
Luo Juan,Yu Yao,Wang Ke,He Sizhe,Wang Longjie,Liang Fangfang,Chin Jason W.,Tang Shan
Catalytic mechanism-based, light-activated traps have recently been developed to identify the substrates of cysteine or serine hydrolases. These traps are hydrolase mutants whose catalytic cysteine or serine are replaced with genetically encoded 2,3-diaminopropionic acid (DAP). DAP-containing hydrolases specifically capture the transient thioester- or ester-linked acyl–enzyme intermediates resulting from the first step of the proteolytic reaction as their stable amide analogs. The trapped substrate fragments allow the downstream identification of hydrolase substrates by mass spectrometry and immunoblotting. In this protocol, we provide a detailed step-by-step guide for substrate capture and identification of the peptidase domain of the large tegument protein deneddylase (UL36 USP ) from human herpesvirus 1, both in mammalian cell lysate and live mammalian cells. Four procedures are included: Procedure 1, DAP-mediated substrate trapping in mammalian cell lysate (~8 d); Procedure 2, DAP-mediated substrate trapping in adherent mammalian cells (~6 d); Procedure 3, DAP-mediated substrate trapping in suspension mammalian cells (~5 d); and Procedure 4, substrate identification and validation (~12–13 d). Basic skills to perform protein expression in bacteria or mammalian cells, affinity enrichment and proteomic analysis are required to implement the protocol. This protocol will be a practical guide for identifying substrates of serine or cysteine hydrolases either in a complex mixture, where genetic manipulation is challenging, or in live cells such as bacteria, yeasts and mammalian cells.
研究领域:
生物化学
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分子生物学
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细胞生物学
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病毒学
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蛋白质组学
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产品关联
DOI
PubMed
高影响因子
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