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Gene expression profile favoring phenotypic reversion: a clue for mechanism of tumor suppression by NF-IL6 3'UTR

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cell Research 2003年第6期13卷 509-514页

作者： DING GAN LIU, QIU HONG JIANG, YUN Yl WEI, LI SUN, BEI BEI FU, Fu KUN ZHAO, QIONG ZHOU ’ state key laboratory of molecular biology and key laboratory of Proteomics, institute of biochemistryand cell biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai200031, china. E-mail: dgliu@*** state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. dgliu@***

Transfection of cDNA in 3’untranslated region of human nuclear factor for interleukin-6 (NF-IL6 3’UTR) induced tumor suppression in a human hepatoma cell line. cDNA array analysis was used to reveal changes in gene expression profile leading to tumor suppression The results indicate that this suppression was not due to activation of dsRNA-dependent protein kinase, nor to inactivation of oncogenes; rather, all the changes in expression of known genes, induced by NF-IL6 3’UTR cDNA may be ascribed to the suppression of cellular malignancy. Therefore, our results imply that this 3’untranslated region may have played role of a regulator of gene expression profile.

关键词： cDNA array NF-IL6 hepatoma 3 ’UTR tumor suppression.

Norepinephrine transporter (NET) is expressed in cardiac sympathetic ganglia of adult rat

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cell Research 2001年第4期11卷 317-320页

作者： LI HE, SUN KAI MA , XIAO PING HU , GOU YUAN ZHANG , JIAN FEI institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, china Cardiovascular Division of Changzheng Hospital, Shang Cardiovascular Division of Changzheng Hospital Shanghai 200003 China

The sympathetic nervous system plays a cardinal role in regulating cardiac function through releasing the neurotransmitter norepinephrine (NE). In comparison with ceniral nervous system, the molecular mechanism of NE uptake in myocardium is not clear. In present study we proved that in rat the CNS type of NE transporter (NET) was also expressed in middle cervical-stellate ganglion complex (MC-SG complex) which is considered to control the activity of heart, but not expressed in myocardium. The results also showed that NET expression level in right ganglion was significantly higher than in the left, rendering the greater capacity of NE uptake in right ventricle, a fact which may contribute to the maintenance of right ventricular function under pathologic state.

关键词： Norepinephrine transporter gene expression cardiac sympathetic ganglion, rat.

The interaction between the human β-globin locus control region and nuclear matrix

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cell Research 2002年第Z2期 411-416页

作者： SHU BING ZHANG, Ruo LAN QIANstate key laboratory of molecular biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, 320 Yueyang Road, shanghai 200031, china

Our previous study showed that hydroxyurea (Hu) could induce HEL cells to express humanβ-globin gene. However the molecular mechanisms by which the expression of β-globin gene is activated and regulated are poorly understood. Here we show that the binding patterns between the core DNA sequences (HS2 core sequence -10681- -10971 bp , HS3 core sequence -14991- -14716 bp and HS4 core sequence -18586- -18306 bp) of DNase I hypersensitive sites in the human β-globin LCR and nuclear matrix proteins isolated from Hu induced and uninduced HEL cells are quite different. Results demonstrated that nuclear matrix proteins might play important roles in regulating the expression of humanβ-like globin genes through their interaction with HSs (HS2,HS3 and HS4 core sequences) in the LCR. Moreover, the results obtained from the in vitro DNA-matrix binding assay showed that the core DNA sequences of DNase I hypersensitive sites (HS2, HS3 and HS4) were unable to bind to the nuclear matrix isolated from uninduced HEL cel

关键词： human β-globin LCR nuclear matrix proteins nuclear matrix HEL cells.

Computational Identification of Novel Family Members of MicroRNA Genes in Arabidopsis thaliana and Oryza sativa

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Acta Biochimica et Biophysica Sinica 2005年第2期37卷 7+76-87页

作者： Yang LI Wei LI You-Xin JIN state key laboratory of molecular biology,institute of biochemistry and cell biology.shanghai institutes for biological sciences,chinese academy of sciences.shanghai 200031.china state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

MicroRNAs(miRNAs)are a class of endogenous small RNAs that play important regulatoryroles in both animals and plants,miRNA genes have been intensively studied in animals,but not in *** study,we adopted a homology search approach to identify homologs of previously validated plantmiRNAs in Arabidopsis thaliana and Oryza *** identified 20 potential miRNA genes in Arabidopsisand 40 in ***,providing a relatively complete enumeration of family members for these miRNAs *** addition,a greater number of Arabidopsis miRNAs(MIR168,MIR159 and MIR172)were found tobe conserved in *** the novel homologs,most of the miRNAs have closely related fellow miRNAs andthe number of paralogs varies in the different miRNA ***,a probable functional segmenthighly conserved on the elongated stem of pre-miRNA fold-backs of MIR319 and MIR159 family *** results support a model of variegated miRNA regulation in plants,in which miRNAs withdifferent functional elements on their pre-miRNA fold-backs can differ in their function or regulation,andclosely related miRNAs can be diverse in their specificity or competence to downregulate target *** that the sophisticated regulation of miRNAs can achieve complex biological effects through qualita-tive and quantitative modulation of gene expression profiles in plants.

关键词： microRNA post-transcriptional gene silencing rice Arabidopsis

Get effective polyclonal antisera in one month

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cell Research 2002年第2期12卷 157-160页

作者： YUAN XIN HU, Ju YUAN QUO, Lu SHEN, YAN CHEN, Zu CHUAN ZHANG, YONG LIAN ZHANGstate key laboratory of molecular biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, 320 Yue Yang Road, 1state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1state key laboratory of molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China

According to the traditional immunization procedure, after the first injection of the sample A (emulsion of aimed antigen and Freund’s complete adjuvant) to immunize rabbit, successive injections of the sample B (emulsion of aimed antigen and Freund’s incomplete adjuvant) were followed every 2-4 weeks. In general, high titer of the corresponding polyclonal antisera will be observed after 4-5 injections of sample B in 3-4 months. This report presents a simply modified procedure that was able to stimulate the antisera formation in one month and achieve enough avidity to satisfy either Western blot or immunohistochemistry analysis. It just applied an additional injection of the sample A to the rabbit at the 3rd day after the primary immunization injection. You could gain the high titer of the antisera right after the first sample B injection in one month. This method has produced the desired results in three different recombinant antigens with different molecular weight (5.9 KD-55 KD) expressed from prokary

关键词： Polyclonal antisera preparation Modified immunization method.

Identification of glutamate transporters and receptors in mouse testis

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Acta Pharmacologica Sinica 2004年第03期25卷 112-117页

作者： Jia-hua HU,Na YANG2,Ying-hua MA,Jie JIANG,Jin-fu ZHANG3,Jian FEI4,Li-he GUO4 laboratory of molecular cell biology,institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031 3Department of Urology,Tongji Hospital,Tongji University,shanghai 200065,china laboratory of molecular cell biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological Sciences Chinese Academy of Sciences Department of Urology Tongji Hospital Tongji University

AIM: To investigate the presence of glutamate transporters and receptors in mouse testis. METHODS: Glutamate uptake analysis was performed to study the function of glutamate transporters in mouse testis. Comparative RT-PCR technique and sequencing analysis were used to study the expression of glutamate receptors and transport- ers in mouse testis. RESULTS: Mouse testis possessed glutamate uptake capacity with sodium-dependence. Vmax value of glutamate uptake was (1.60 ± 0.21) pmol/min per mg protein and Km value of glutamate uptake was (11.0±1.6) μmol/L in mouse testis according to saturation analysis. Furthermore, the uptake activity could be inhibited by DHK (GLT1 selective inhibitor) and THA (glutamate uptake inhibitor). In addition, RT-PCR results revealed that glutamate transporters (GLT1 and EAAC1) and ionotropic glutamate receptors (NR1, NR2B, GluR6 and KA2) were expressed in mouse testis. CONCLUSION: Glutamate transporters and receptors do exist in mouse testis.

关键词： glutamates receptors amino acid transport systems testis uptake

Analysis of the Interaction between hPFTAIRE1 and PLZF in a Yeast Two-hybrid System

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Acta Biochimica et Biophysica Sinica 2006年第3期38卷 164-170页

作者： Yan-Kun GAO~# Mei JIANG~# Tao YANG Jiang-Ye CHEN~* state key laboratory of molecular biology,institute of biochemistry,and cell biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china statekeylaboratoryofmolecularbiology InstituteofBiochemistryandCellBiologyShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesShanghai200031China

hPFTAIRE1 is a Cdc2-related kinase family *** search its substrates and regulatoryproteins,hPFTAIRE1 was fused to LexA and used as a bait to screen a human brain LexA two-hybrid *** this screening,seven hPFTAIRE1 interacting proteins,including promyelocytic leukemia zinc finger (PLZF),were *** interaction between PLZF and hPFTAIRE1 was confirmed by β-galactosidase assay andLeu growth *** encodes a transcription factor belonging to the POZ/BTB domain and Krüppelzinc finger (POK) *** highly conserved POZ/BTB domain plays a critical role in *** deleted the POZ/BTB and Krüppel zinc finger domains,respectively,and observed the inter-action between hPFTAIRE1 and truncated PLZFs by liquid β-galactosidase activity assay.A weak interactionwas detected between hPFTAIRE1 and *** also observed the interaction between PLZF and anotherCdc2-related kinase,PCTAIRE1.A similar result was *** interaction between PLZF and hPFTAIRE1or PCTAIRE1 was confirmed by co-immunoprecipitation assay in a yeast *** is a phosphoproteinand plays multiple roles during cell *** results suggest that hPFTAIRE1 and PCTAIRE1 may playimportant roles in the functional regulation of PLZF.

关键词： hPFTAIRE1 PLZF PCTIARE1 yeast two-hybrid interaction

Direct transfer of A20 gene into pancreas protected mice from streptozotocin-induced diabetes

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Acta Pharmacologica Sinica 2004年第6期25卷 19-24页

作者： Lu-yang YU, Bo LIN, Zhen-lin ZHANG2, Li-he GUO3 instituteof biochemistryand cell biology, shanghai institutes for biological sciences, chinese academy ofsciences, shanghai 200031 Center for Preventing and Treating Osteoporosis, Osteoporosis Research Unit, 2 shanghai JiaoTong UniversityAffiliated Sixth People’s Hospital, shanghai 200233, china institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Center for Preventing and Treating Osteoporosis Osteoporosis Research Unit Shanghai JiaoTong University Affiliated Sixth People’s Hospi

AIM: To investigate the efficiency of transfer of A20 gene into pancreas against STZ-induced diabetes. METHODS: PVP-plasmid mixture was directly transferred into the pancreatic parenchyma 2 d before STZ injection. The uptake of plasmid pcDNA3-LacZ or pcDNA3-A20 was detected by PCR and the expression of LacZ was confirmed by histological analysis with X-gal. A20 expression in the pancreas of pcDNA3-A20 transgenic mice was measured by RT-PCR and Western blots. Urine amylase, NO generation, and histological examination were examined. RESULTS: Injection of PVP-plasmid mixture directly into the pancreatic parenchyma increased urine amylase concentration 16 h after operation and reversed it to nearly normal 36 h later. On d 33 LacZ expression could be found in spleen, duodenum, and islets. The development of diabetes was prevented by direct A20 gene transferring into the pancreas and A20-mediated protection was correlated with suppression of NO production. The insulitis was ameliorated in A20-treated mice. CONCLUSION: Injection of PVP-plasmid mixture directly into the pancreatic parenchyma led to target gene expression in islets. Direct transfer of A20 gene into the pancreas protected mice from STZ-induced diabetes.

关键词： adenoviridae streptozocin insulin-dependent diabetes mellitus gene therapy nitric oxide glucose amylases reversed transcriptase polymerase chain reaction Western blotting

Characterization of ATPase Activity of Recombinant Human Pif1

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Acta Biochimica et Biophysica Sinica 2006年第5期38卷 335-341页

作者： Yu HUANG Deng-Hong ZHANG Jin-Qiu ZHOU Max-Planck Junior Research Group at the state key laboratory of molecular biology,institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china Max-Planck Junior Research Group at the state key laboratory of molecular biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China

Saccharomyces cerevisiae Piflp helicase is the founding member of the Pifl subfamily that isconserved from yeast to *** potential human homolog of the yeast PIFI gene has been cloned fromthe cDNA library of the Hek293 cell ***,we described a purification procedure of glutathione S-transferase（GST）-fused N terminal truncated human Pifl protein（hPif1ΔN）from yeast and characterizedthe enzymatic kinetics of its ATP hydrolysis *** ATPase activity of human Pif1 is dependent ondivalent cation,such as Mg2+,Ca2+and single-stranded *** for ATP for the ATPase activity isapproximately 200 μ*** the ATPase activity is essential for hPifl’s helicase activity,these results willfacilitate the further investigation on hPif1.

关键词： Pifl helicase telomere telomerase ATPase