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Interaction of MHV RNA with mtHSP70 and m-aconitase

MHV RNA 与 mtHSP70 和 m-乌头酸酶的相互作用

基本信息

批准号：
7159356
负责人：
JULIAN L LEIBOWITZ
金额：
$ 24.14万
依托单位：
TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-07-01 至 2008-12-31
项目状态：
已结题

项目摘要

We have recently identified four proteins which interact with a protein binding element [3'(+)42 element] contained within the last 42 nucleotides of the mouse hepatitis virus genome. In this application we propose to examine the interaction of these four proteins, mitochondrial HSP70 (mtHSP70), mitochondrial aconitase (m-aconitase), HSP60, and HSP40, with MHV RNA. We will employ fluorescence resonance energy trar_sfer (FRET), immuno-electron microscopy, and cell permeant cross-linking studies to verify that the interaction of these proteins with MHV RNA takes place in intact cells. We will undertake a series of biochemical studies to determine if the MHV RNA interacts with mtHSP70, m-aconitase, and HSP60 prior to the importation of these proteins into mitochondria, or alternatively, if these MHV RNA interacting proteins are exported from mitochondria. We will also determine if mitochondrial function is altered during MHV infection. To better characterize the structural basis for these interactions we will perform a series of experiments to map the residues of m-aconitase, mtHSP70, HSP60, and HSP40 which are in contact with the MHV 3'(+)42 protein binding element. We will subsequently carry out a series of mutagenesis experiments to map residues of these proteins required for RNA binding. A second line of mutagenesis experiments will more precisely define the sequence requirements of the RNA for protein binding. We have determined that these proteins interact during binding, thus dominant negative mutants will be particularly sought. We have determined that mtHSP70 is tyrosine phosphorylated, and that the phosphorylation state of the MHV 3'(+)42 binding proteins regulates their binding the MHV 3'(+)42 element. We will perform a series of pharmacologic and genetic manipulations to investigate the role of tyrosine phosphorylation in regulating the interaction of these proteins with MHV RNA. The functional significance of the interaction of mtHSP70, m-aconitase, HSP60, and HSP40 with MHV RNA on virus replication will be examined in a series of experiments employing mutational and over-expression strategies.

我们最近发现了四种与蛋白质结合元件[3'(+)42元件]相互作用的蛋白质

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mouse hepatitis virus stem-loop 4 functions as a spacer element required to drive subgenomic RNA synthesis.

小鼠肝炎病毒茎环 4 充当驱动亚基因组 RNA 合成所需的间隔元件。

DOI：
10.1128/jvi.05092-11
发表时间：
2011
期刊：
Journal of virology
影响因子：
5.4
作者：
Yang,Dong;Liu,Pinghua;Giedroc,DavidP;Leibowitz,Julian
通讯作者：
Leibowitz,Julian

In vitro assembled, recombinant infectious bronchitis viruses demonstrate that the 5a open reading frame is not essential for replication.

DOI：
10.1016/j.virol.2004.10.045
发表时间：
2005-02-05
期刊：
Virology
影响因子：
3.7
作者：
Youn S;Leibowitz JL;Collisson EW
通讯作者：
Collisson EW

SHAPE analysis of the RNA secondary structure of the Mouse Hepatitis Virus 5' untranslated region and N-terminal nsp1 coding sequences.

DOI：
10.1016/j.virol.2014.11.001
发表时间：
2015-01-15
期刊：
VIROLOGY
影响因子：
3.7
作者：
Yang, Dong;Liu, Pinghua;Wudeck, Elyse V.;Giedroc, David P.;Leibowitz, Julian L.
通讯作者：
Leibowitz, Julian L.

The virion N protein of infectious bronchitis virus is more phosphorylated than the N protein from infected cell lysates.