Cdc42在甲型流感病毒NA蛋白修饰及转运中的作用
[Abstract]:Influenza virus neuraminidase (NA) has enzyme activity and plays an important role in virus invasion and release. According to its characteristics, NA has become the most suitable target for the design of anti-influenza drugs. At present, although studies continue to improve the NA inhibitors, influenza virus strains are still rapidly developing drug resistance to these drugs. Therefore, it is urgent to develop new anti-influenza drugs and find new targets. In host cells, many host proteins play an important role in the replication of influenza viruses. Among them, Cdc42,RhoA and Rac1, three important family members of small G protein RhoGTPases, are involved in many cell signal transduction pathways and participate in many cellular biological activities as molecular switches. More and more studies show that they are involved in the regulation of intracellular transport. However, whether they play a role in intracellular transport of NA protein and which of them is involved has not been reported. This paper studies this problem. We first observed the localization of NA and NA (H274Y) mutants in cells and detected the decrease of surface enzyme activity of NA (H274Y) mutants, and established a system of intracellular transport of NA proteins. Then, the cells transfected with NA were treated with Toxin B, which confirmed that Rho GTPases family members (Cdc42, RhoA and Rac1) promoted the intracellular transport of NA protein. Then we observed the co-localization of Cdc42,RhoA and Rac1 and their continuous activation and inhibition with NA protein, as well as the drug C3Transferase treatment. It was proved that Cdc42 played a promoting role. Our exogenous transfer into Cdc42, a specific GTP activator protein, (GAP)-ARHGAP21, inhibited the activity of Cdc42 in host cells, further confirming the role of endogenous Cdc42. We also studied the interaction between Cdc42 and NA protein. GST-pulldown test showed that there was no direct interaction between Cdc42 and NA. TGN and GM130 were used to label Golgi body respectively. Confocal test showed that Cdc42 affected the transport of NA protein from Golgi body to cytoplasm and cell membrane. Subsequently, the Cytochalasin D treatment test revealed that Cdc42 affected the transport of NA protein through the regulation of actin. In addition, we constructed a series of NA deletion mutants for NA signal peptide region. We found that the absence of more than 10 amino acids in the transmembrane region of NA resulted in no expression of NA protein. We studied the protein transport of the mutant with NA deletion of 11 amino acid. The results showed that the localization in the cell had not changed. These results suggest that Cdc42 plays an important role in the modification and transport of NA protein of influenza A virus. The mechanism of Cdc42 in NA protein transport needs further study.
【学位授予单位】:福建农林大学
【学位级别】:硕士
【学位授予年份】:2011
【分类号】:R373.13
【共引文献】
相关期刊论文 前10条
1 ;Diagnosis and Prevention of Swine Influenza[J];Animal Husbandry and Feed Science;2009年Z2期
2 王新卫;詹爱军;;禽流感病毒的宿主特异性和致病性分子基础[J];长江大学学报(自科版)农学卷;2006年01期
3 郭元吉;;正确面对禽流感病毒对人类健康的威胁[J];传染病信息;2006年01期
4 仇华吉,童光志;猪流感的再认识[J];动物医学进展;2000年03期
5 董婕;禽流感病毒感染人的机理研究[J];公共卫生与预防医学;2005年04期
6 Masoud Soltanialvar;Reza Goodarzi;Farshad Akbarnejad;;Genetic analysis of polymerase complex(PA,PB1 and PB2) genes of H9N2 avian influenza viruses from Iran(1999 to 2009)[J];Asian Pacific Journal of Tropical Biomedicine;2012年11期
7 齐慧川;张yN;胡敏;;实验性正畸牙移动过程中EphB4/ephrinB2促成骨作用的研究进展[J];吉林大学学报(医学版);2013年06期
8 Jin-qian Zhang;Li-cheng Zhang;Na Ren;Ming Zhang;Li-min Guo;Xing-wang Li;Jun Cheng;;Factors Associated with Death Due to 2009 Influenza A (H1N1) Virus Infection and Acute Respiratory Distress Syndrome in Beijing, 2009-2011[J];Infection International(Electronic Edition);2012年04期
9 李凤茹;江华;;Rho蛋白与恶性肿瘤相关性的研究进展[J];国际消化病杂志;2014年01期
10 谭伟;徐倩;谢芝勋;;禽流感病毒研究概述[J];基因组学与应用生物学;2014年01期
相关会议论文 前5条
1 李敏;向华;严悌昆;;猪流感的流行及其公共卫生学意义[A];提高全民科学素质、建设创新型国家——2006中国科协年会论文集[C];2006年
2 关启刚;曾定尹;孙喜琢;周旭晨;程颖;苗志林;何学志;韩凤桐;张利;;通心络对IL-1β介导的小型猪冠状动脉内膜增殖及5-HT诱发的冠状动脉痉挛的抑制作用[A];首届中西医血管病学大会论文汇编[C];2013年
3 熊新贵;陈疆;梁清华;杨波;吴丹;;类风湿关节炎患者关节滑膜组织蛋白质组学研究[A];第十一次中国中西医结合实验医学学术研讨会论文汇编[C];2013年
4 Ping Huang;Yuansheng Xu;Hanzhong Ni;Jing Zhong;Xin Zhang;Songnuan Tan;De Wu;Baoyun Qiu;Dawei Guan;Miaoheng Wen;Jin Yan;Yonghui Zhang;;Linear B-cell epitope mapping of neuraminidases of the 2009 A H1N1 viruses based on immunoinformatics[A];2011广东省预防医学会学术年会资料汇编[C];2011年
5 李铁钢;Chuanxi Fu;Biao Di;Jibin Wu;Zhicong Yang;Yulin Wang;Meixia Li;Jianyun Lu;Yiyun Chen;Enjie Lu;Jinmei Geng;Wensui Hu;Zhiqiang Dong;Meng-feng Li;Bo-Jian Zheng;Kai-yuanCao;Ming Wang;;A Two-Year Surveillance of 2009 Pandemic Influenza A (H1N1) in Guangzhou,China:From Pandemic to Seasonal Influenza?[A];2012广东省预防医学会学术年会资料汇编[C];2013年
相关博士学位论文 前10条
1 张文韬;基于量子点的禽流感病毒检测新方法探索[D];武汉大学;2010年
2 贾永清;表达禽流感病毒HA基因重组禽痘病毒的构建及其免疫效力的研究[D];东北农业大学;2000年
3 刘明;H5和H7亚型禽流感分子防制技术的研究[D];中国农业科学院;2000年
4 乔传玲;表达禽流感病毒HA-NA、HA-NP及NP基因重组禽痘病毒的构建及其免疫效力的研究[D];中国农业科学院;2001年
5 杨建德;马流感病毒青海株相关特性研究及其基因组分析[D];东北农业大学;2003年
6 马文军;禽流感病毒和鲍纳病毒反向遗传系统的研究[D];中国农业科学院;2003年
7 张云;宠物鸟H3N8亚型流感病毒的鉴定及利用反义遗传学技术产生重组病毒[D];中国农业科学院;2003年
8 陈素娟;用不同鸡痘病毒载体构建单表达或双表达抗H5和H9亚型禽流感的重组疫苗及其免疫效力[D];扬州大学;2005年
9 杨彩然;禽源H3、H4亚型流感病毒的序列分析及对鸡的致病性研究[D];内蒙古农业大学;2005年
10 贾雷立;禽流感病毒分离、复合多表位重组体构建及实验免疫研究[D];吉林大学;2006年
相关硕士学位论文 前10条
1 郑毅;利用酵母双杂交技术研究流感病毒NS2蛋白与宿主蛋白的相互作用[D];华中农业大学;2010年
2 朱晓林;三株不同亚型SIV HA基因与NA基因克隆与序列分析[D];山东农业大学;2010年
3 崔佳莹;H5N1亚型禽流感病毒单克隆抗体的制备与血凝素HA基因的体外高效表达和纯化[D];中国农业科学院;2011年
4 王耿燕;一株禽H1N1流感病毒血凝素蛋白的纯化、鉴定和结构分析[D];汕头大学;2011年
5 侯君;大连市流感病毒血凝素基因遗传特性研究[D];大连医科大学;2011年
6 邓国华;禽流感病毒分离株A/Chicken/Xinjiang/1/96(H14N5)全基因克隆及其序列分析[D];东北农业大学;2000年
7 张梅;河北省人群中猪型(H1N1)流感病毒血清学调查的研究[D];河北农业大学;2001年
8 徐克敏;华南地区鸭源性H9亚型流感病毒的基因特性研究[D];汕头大学;2002年
9 焦培荣;一株H5N1亚型禽流感病毒的生物学特性及其感染性克隆的研究[D];内蒙古农业大学;2004年
10 陈君彦;H1N1亚型猪流感病毒中国分离株的分子遗传衍化的研究[D];内蒙古农业大学;2004年
,本文编号:2274442
本文链接:https://www.wllwen.com/xiyixuelunwen/2274442.html