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| 1 | Expression and subcellular localization of mechano-growth factor in osteoblasts under mechanical stretch显示文摘Mechano-growth factor (MGF) is a stretch sensitive factor in myocytes, and it might also be produced by other mechanocytes under mechanical stimulation. In this study, both the mRNA and protein expression of MGF were detected in stretched osteoblasts. Quantitative analysis showed that a cyclic stretching stimulation caused a quick and sharp increase of MGF mRNA and protein expression from a low basal level under no stretch; the mRNA and protein levels respectively peaked in 6 and 12 h to 5 and 5.2 fold over the basal level and returned to normal by 24 h. The subcellular distribution of MGF protein was revealed by immunofluorescence analysis to be restricted to the nucleus. We concluded that cyclic stretching stimulation could induce MGF expression in osteoblasts in a pulsing fashion; and the nuclear distribution of MGF suggested that MGF might act in mechanocytes as an autocrine growth factor. | ZHANG BingBing, XIAN ChengYu, LUO YanFeng & WANG YuanLiang Research Center of Bioinspired Materials Science and Engineering, Bioengineering College, Chongqing University, Chongqing 400030, China | 2009 | Science China(Life Sciences)2009，52，10： | 10 |
| 2 | Physiological roles of mitogen-activated-protein-kinase-activated p38-regulated/activated protein kinase显示文摘Mitogen-activated protein kinases(MAPKs)are a family of proteins that constitute signaling pathways involved in processes that control gene expression,cell division, cell survival,apoptosis,metabolism,differentiation and motility.The MAPK pathways can be divided into conventional and atypical MAPK pathways.The first group converts a signal into a cellular response through a relay of three consecutive phosphorylation events exerted by MAPK kinase kinases,MAPK kinase,and MAPK.Atypical MAPK pathways are not organized into this three-tiered cascade.MAPK that belongs to both conventional and atypical MAPK pathways can phosphorylate both non-protein kinase substrates and other protein kinases.The latter are referred to as MAPK-activated protein kinases.This review focuses on one such MAPK-activated protein kinase,MAPK-activated protein kinase 5(MK5)or p38-regulated/activated protein kinase(PRAK).This protein is highly conserved throughout the animal kingdom and seems to be the target of both conventional and atypical MAPK pathways.Recent findings on the regulation of the activity and subcellular localization,bona fide interaction partners and physiological roles of MK5/PRAK are discussed. | Sergiy Kostenko Gianina Dumitriu Kari Jenssen Lgreid Ugo Moens | 2011 | World Journal of Biological Chemistry2011，2，5： | 8 |
| 3 | A transient expression system in soybean mesophyll protoplasts reveals the formation of cytoplasmic GmCRY1 photobody-like structures显示文摘Soybean(Glycine max(L.) Merr.), grown for its plant oils and proteins, is one of the most important crops throughout the world.Generating stable and heritable transgenic soybeans is relatively inefficient;therefore, there is an urgent need for a simple and high-efficient transient transformation method by which to enable the investigation of gene functions in soybeans, which will facilitate the elucidation and improvement of the molecular mechanisms regulating the associated agronomic traits. We established a system of transient expression in soybean mesophyll protoplasts and obtained a high level of protoplast transfection efficiency(up to 83.5%). The subcellular activity of the protoplasts was well preserved, as demonstrated by the dynamic formation of GmCRY nucleus photobodies(NPs) and/or cytoplasmic photobody-like structures(CPs) in response to blue light.In addition, we showed that GmCRY1b CPs colocalized with GmCOP1b, a co-ortholog of Arabidopsis thaliana CONSTITUTIVE PHOTOMORPHOGENIC 1(COP1), which provided new insight into the potential roles of GmCRY1s in the cytoplasm. | Lu Xiong Cong Li Hongyu Li Xiangguang Lyu Tao Zhao Jun Liu Zecheng Zuo Bin Liu | 2019 | Science China(Life Sciences)2019，62，8： | 6 |
| 4 | Rice Heavy Metal P-type ATPase OsHMA6 Is Likely a Copper Efflux Protein显示文摘P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phylogenetic tree analysis showed that OsHMA6 belonged to the Cu/Ag subgroup of the HMA family and had a close evolutionary relationship with OsHMA9.Amino acid sequence alignment showed 82.78%consistency between OsHMA6 and OsHMA9.OsHMA6 expressed in all organs at different growth stages,including spikelet,and abundant in leaf blades,however,OsHMA9 most strongly expressed in roots,but very low in spikelet.Excessive Cu^2+can up-regulate the expression of OsHMA6 and OsHMA9 in rice seedlings.The heterologous expression in yeast showed that OsHMA6 can significantly rescue the growth of yeast strain CM52 when supplied with 3 or 6 mmol/L Cu^2+.Compared with the empty vector pYES2,the Cu concentration in OsHMA6-pYES2 decreased by 23.4%and 30.3%under 3 or 6 mmol/L Cu2+,respectively.Subcellular localization revealed that OsHMA6 was located in the plasma membrane.These results suggested that OsHMA6,similar to OsHMA9,is likely a copper efflux protein located in the plasma membrane. | ZOU Wenli LI Chang ZHU Yajun CHEN Jingguang HE Haohua YE Guoyou | 2020 | Rice science2020，27，2： | 2 |
| 5 | Characterization and Expression Analysis of Protein Kinase C Gene from Dunaliella salina显示文摘Protein kinase C (PKC) has a crucial role in signal transduction for a variety of biologically active substances which activate cellular functions and proliferation. We previously isolated the full-length PKC gene from Dunaliella salina (DsPKC) using rapid amplification of cDNA ends (RACE) and RT-PCR methods. And we submitted the mRNA sequence of DsPKC gene to NCBI (Genbank No. JN625213). In the present paper, the DsPKC gene open reading frame obtained by PCR was cloned into pGS-21a vector and transformed into Escherichia coli to generate the fusion protein. Bioinformatics analysis revealed that DsPKC gene was a member of serine/threonine kinase with two conserved domains and highly conserved motifs. The DsPKC was highly expressed upon induction with isopropyl-β-d-thiogalactoside (IPTG) at a final concentration of 0.2 mmol L 1 at 37℃. Under salt stress, the fu- sion protein Green Fluorescent Protein (GFP)-DsPKC was transferred from the cytoplasm to the cell membrane. The expression pat- tern of DsPKC gene was analyzed using real-time quantitative PCR, and indicated that DsPKC gene was up-regulated by 3.0 mol L 1 NaCl at 12 h, which was significantly higher than in control values (P < 0.05). These results suggest that the DsPKC gene plays an important role in response to salt stress in D. salina. | CONG Yuting MA Yuexin WANG Yuan LIU Yiqiong CHAI Xiaojie | 2019 | Journal of Ocean University of China2019，18，4： | 2 |
| 6 | Identification and characterization of BoPUB3: a novel interaction protein with S-locus receptor kinase in Brassica oleracea L.显示文摘Armadillo repeat containing 1 (ARC1) is phosphorylated by S-locus receptor kinase (SRK) and functions as a positive regulator in self-incompatibility response of Brassica. However, ARC1 only causes partial breakdown of the self-incompatibility response, and other SRK downstream factors may also participate in the self-incompatibility signaling pathway. In the present study, to search for SRK downstream targets, a plant U-box protein 3 (BoPUB3) was identified from the stigma of Brassica oleracea L. BoPUB3 was highly expressed in the stigma, and its expression was increased with the stigma development and reached to the highest level in the mature-stage stigma. BoPUB3, a 76.8-kDa protein with 697 amino acids, is a member of the PUB-ARM family and contains three domain characteristics of BoARC1, including a U-box N-terminal domain, a U-box motif, and a C-terminal arm repeat domain. The phylogenic tree showed that BoPUB3 was close to BoARC1. The synteny analysis revealed that B. oleracea chromosomal region containing BoPUB3 had high synteny with the Arabidopsis thaliana chromosomal region containing AtPUB3 (At3G54790). In addition, the subcellular localization analysis showed that BoPUB3 primarily localized in the plasma membrane and also in the cytoplasm. The combination of the yeast two-hybrid and in vitro binding assay showed that both BoPUB3 and BoARC1 could interact with SRK kinase domain, and SRK showed much higher level of β-galactosidase activity in its interaction with BoPUB3 than with BoARC1. These results implied that BoPUB3 is a novel interactor with SRK, which lays a basis for further research on whether PUB3 participates in the self-incompatibility signaling pathway. | Songmei Shi Qiguo Gao Tonghong Zuo Zhenze Lei Quanming Pu Yukui Wang Guixi Liu Xinhua He Xuesong Ren Liquan Zhu | 2019 | Acta Biochimica et Biophysica Sinica2019，51，7： | 2 |
| 7 | Protein trafficking in plant cells:tools and markers显示文摘Eukaryotic cells consist of numerous membrane-bound organelles,which compartmentalize cellular materials to fulfil a variety of vital functions.In the post-genomic era,it is widely recognized that identification of the subcellular organelle localization and transport mechanisms of the encoded proteins are necessary for a fundamental understanding of their biological functions and theorganization of cellular activity.Multiple experimental approaches are now available to determine the subcellular localizations and dynamics of proteins.In this review,we provide an overview of the current methods and organelle markers for protein subcellular localization and trafficking studies in plants,with a focus on the organelles of the endomembrane system.We also discuss the limitations of each method in terms of protein colocalization studies. | Dongmei Zhu Mengdi Zhang Caiji Gao Jinbo Shen | 2020 | Science China(Life Sciences)2020，63，3： | 2 |
| 8 | 水稻OsWRKY78与GFP融合基因的拟南芥转化及亚细胞定位(英文)显示文摘[目的]确定OsWRKY78蛋白在植物中的定位。[方法]根据GenBank数据库中OsWRKY78全序列设计引物,进行OsWRKY78的RT-PCR扩增,克隆了OsWRKY78基因,将该片段与带绿色荧光蛋白(GFP) 基因的质粒载体pBinGFP重组,并对重组载体进行菌液PCR和酶切验证,最后利用农杆菌介导的花蕾浸泡法将重组载体转化到拟南芥中,对其亚细胞定为进行研究。[结果]试验克隆得到了pBinGFP-OsWRKY78重组载体,经菌落PCR与酶切检测表明构建的表达载体正确,其转化到拟南芥中后得到了转基因植株,荧光显微镜检测结果表明,OsWRKY78基因表达产物主要定位在细胞核中。[结论]该研究结果为深入研究OsWRKY78基因的功能及其在相关信号传导中的作用奠定了基础,也为进一步研究OsWRKY78基因与褐飞虱之间的关系提供了理论依据。 | 刘顺枝 张美 唐馨 王小兰 | 2012 | Agricultural Science & Technology2012，13，7： | 1 |
| 9 | Subcellular Localization of Large Yellow Croaker(Larimichthys crocea) TLR21 and Expression Profiling of Its Gene in Immune Response显示文摘Toll-like receptor 21(TLR21) is a non-mammalian type TLR, and plays an important role in innate immune response in fish. In this paper, the full-length cDNA sequence of TLR21 gene was identified and characterized from large yellow croaker, Larimichthys crocea and was termed as LcTLR21. It consists of 3365 bp, including a 5'-terminal untranslated region(UTR) of 97 bp, a 3'-terminal UTR of 331 bp, and an open reading frame(ORF) of 2937 bp encoding a polypeptide of 978 amino acid residues. The deduced LcTLR21 contains a signal peptide domain at N-terminal, 12 leucine-rich repeats(LRRs) at the extracellular region, a transmembrane domain and a cytoplasmic toll-interleukin-1 receptor(TIR) domain at the C-terminal. Subcellular localization analysis revealed that the LcTLR21-GFP was constitutively expressed in cytoplasm. Tissue expression analysis indicated that LcTLR21 gene broadly expressed in most of the examined tissues, with the most predominant abundance in spleen, followed by head-kidney and liver, while the weakest expression was detected in brain. The expression level of LcTLR21 after LPS, poly I:C and Vibrio parahaemolyticus challenges was investigated in spleen, head-kidney and liver. LcTLR21 gene transcripts increased significantly in all examined tissues after the challenges, and the highest expression level was detected in liver at 24 h after poly I:C stimulation(P <0.05), suggesting that LcTLR21 might play a crucial role in fish resistance to viral and bacterial infections. | SUN Qingxue FAN Zejun YAO Cuiluan | 2018 | Journal of Ocean University of China2018，17，2： | 1 |
| 10 | Cloning and characterization of the PtVIP1 gene in Populus显示文摘The VirE2-interaction protein 1(VIP1)serves as a regulator of mitogen-activated protein kinase 3(MPK3)-mediated stress gene modulation under biotic stress,which in turn activates the MPK3 pathway in Arabidopsis.The mode of action of the VIP1 protein in Populus in response to biotic stress remains unknown.In this study,we cloned the full-length cDNA of the PtVIP1 gene from Populus trichocarpa(accession number of GenBank:KY793105).The VIP1 protein harboured a conserved bZIP(basic leucine zipper)domain located in the C-terminus.The VIP1 subcellular localization assay indicated that the VIP1 protein was present in the cytoplasm and nucleus under normal conditions,and that an increase in the amount of the protein in the nucleus occurred after treatment with flg22,the elicitor-active epitope of flagellin which triggers the innate immune response in plants.Transgenic Populus plants overexpressing VIP1 genes(PtVIP1 of Populus;or AtVIP1 of Arabidopsis,as positive control)were generated to investigate the role of VIP1 in vivo.The expression of poplar pathogenesis-related protein 1(PR1)genes was upregulated in transgenic-PtVIP1 or AtVIP1 poplar plants.The transgenic poplar plants overexpressing PtVIP1 or AtVIP1 also showed enhanced resistance to Brenneria salicis infection.These results suggest that the VIP1 protein accumulates in the nucleus in response to biotic stress,and that the pathogen resistance of transgenic VIP1 poplar may be associated with the induced expression of PR1 genes in response to pathogen challenge. | Haoran Wang Weitao Zhang Mingxiu Wang Qiang Cheng | 2019 | Journal of Forestry Research2019，30，6： | 1 |
| 11 | Identification and characterization of Letm1 gene in Toxoplasma gondii显示文摘Toxoplasma gondii is an obligate intracellular protozoan that causes toxoplasmosis.Previous studies have shown that the perturbation of mitochondrial metabolism in T.gondii results in growth deficiency in host cells and lack of virulence in animals.Members of this Letm1 protein family are inner mitochondrial membrane proteins which play a role in potassium and hydrogen ion exchange.Letm1 has not been characterized in T.gondii.In this study,a potential TgLetm1 gene (TgGT1_288400) with Letm1-like protein domain coding sequence was identified in T.gondii.Indirect immunofluorescence assays suggested that TgLetm1 localized to the mitochondria in tachyzoites,as indicated by the colocalization with mitochondrial marker Mitotracker.TgLetm1 was found in the membrane fraction by western blot analysis.To investigate the role of TgLetm1 in T.gondii,we generated a tetracycline-inducible TgLetm1-knock-down mutant.The conditional deletion of TgLetm1 resulted in mitochondrial swelling.Functional studies showed that the conditional deletion of TgLetm1 resulted in growth inhibition,deficiency in invasion and replication,and lack of virulence in mice. | Le Chang Xichen Zhang Pengtao Gong Yuru Wang Boya Du Jianhua Li | 2019 | Acta Biochimica et Biophysica Sinica2019，51，1： | 1 |
| 12 | Expression, purification, and subcellular localization of phospholipase C in Dunaliella salina显示文摘Plants possess effective mechanisms to respond quickly to the external environment. Rapid activation of phosphatidylinositol-specific phospholipase C (PLC) enzymes occurs after a stimulus. The PLC in Dunaliella salina plays important roles in growth and stress responses. However, the molecular basis of PLC action in D. salina remains little understood. To gain insight into the potential biological functions of this enzyme, we cloned a phospholipase C gene from D. salina in a previous study, named DsPLC (GenBank No. KF573428). Here, we present the prokaryotic expression, purification, and characterization of the DsPLC gene. The entire coding region of DsPLC was inserted into an expression vector pET32a, and the DsPLC gene was successfully expressed in Escherichia coli. The DsPLC protein was purified and identified using a polyclonal antibody and western blotting. Expressing DsPLC fused with a green fluorescent protein (GFP) in onion showed that DsPLC-GFP was localized to the intracellular membrane. Quantitative real-time PCR analysis revealed that the relative expression of the DsPLC gene was induced significantly by 3.0-mol/L NaCl at 4 h. Our results support the importance of PLC enzymes in plant defense signaling. This study provides a basis for further functional studies of the DsPLC gene and for additional analysis of the potential roles of PLC enzymes in response to abiotic stress. | CONG Yuting WANG Yuan YUE Jinrong XING Zhenyu GAO Xiangnan CHAI Xiaojie | 2019 | Journal of Oceanology and Limnology2019，37，4： | 0 |
| 13 | Transcriptional dysregulation in neurodegenerative diseases:who tipped the balance of Yin Yang 1 in the brain?显示文摘Yin Yang 1(YY1) is a multi-functional transcription factor that regulates gene expression in a range of cell types,including neurons.It controls neuronal differentiation,as well as neuronal specification and migration during the development of the mammalian nervous system.Besides,YY1 also mediates the transcription of genes that are required for neuronal survival.An impairment of the transcriptional function of YY1 causes neuronal death.This review summarizes recent research findings that unveil the dysfunction of YY1 in multiple neurodegenerative disorders.The expression of disease proteins perturbs the function of YY1 via distinct molecular mechanisms,including recruitment to protein aggregates,protein degradation and aberrant nuclear/cytoplasmic shuttling.Understanding the pathogenic roles of YY1 will further broaden our knowledge of the disease mechanisms in distinct neurodegenerative disorders. | Zhefan Stephen Chen Ho Yin Edwin Chan | 2019 | Neural Regeneration Research2019，14，7： | 0 |
| 14 | Genome-wide identification, phylogenetic and expression analysis of SMAX1-Like genes in rapeseed(Brassica napus L.)显示文摘SMAX1-LIKE(SMXL) family members have played important roles in regulating plant growth and development in Arabidopsis and rice, but few have been studied in Brassica napus(rapeseed). In this study, 31 SMXL genes(BnaSMXL) were identified in rapeseed by whole genome bioinformatics. Detailed information were characterized, including genomic distribution, phylogenetic relationship, evolutionary selection pattern, structural and motif features. Phylogenetic analysis classified BnaSMXL genes into 4 distinct sub-clades representing clear orthologous relationships to their family members in Arabidopsis and rice. Conserved motif analysis indicated that the motif 16 mediating strigolactones(SLs) or karrikins(KARs) signaling was absent in SMXL3-5 genes. Orthologous gene pairs between Arabidopsis and 3 Brassica species were extracted, and 8 SMXL genes in Arabidopsis genome had 16, 16 and 31 corresponding genes in B. oleracea, B. rapa and rapeseed genomes respectively. Evolutionary selection pattern showed that most of SMXL genes had undergone stronger negative selection. BnaSMXLs expression showed diverse patterns in 10 tissues of both vegetative and reproductive organs. Subcellular localization analysis showed that BnaSMXL proteins were exclusively localized in nucleus. | Hongli Yang Xiaokang Li Jinglin Liu Hongfang Liu Wei Hua Ming Zheng | 2018 | Oil Crop Science2018，3，2： | 0 |
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