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The effect of the regulatory mechanism of splicing on influenza C virus replication

剪接调控机制对丙型流感病毒复制的影响

基本信息

批准号：
17590413
负责人：
HONGO Seiji
金额：
$ 2.24万
依托单位：
Yamagata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590413/
关键词：
influenza C virus splicing NS gene regulation of gene expression

项目摘要

We recently established reverse genetics of influenza C virus with seven Pol I plasmids expressing PB2, PB 1, P3, HE, NP, M and NS viral RNA and nine plasmids expressing PB2, PB1, P3, HE, NP, M1, CM2, NS1 and NS2 proteins and succeeded in generation of a recombinant influenza C virus. To determine whether NS gene products of influenza C virus play a role in pre-mRNA splicing, we attempted to obtain the virus, in which Leu 20 was substituted into termination codon to rescue the virus with little expressing of NS1 and NS2. However, the virus was not rescued, which suggested that NS gene products, NS1 and/or NS2, are required for virus replication. In the transient expression system, coexpression of M gene cDNA with NS1 protein showed that splicing rate of M pre-mRNA increased by 30%, suggesting that NS1 may have the activity to enhance splicing. We then investigated whether C-terminus region (63-246 residues) specific for NS1 possesses the functional domain to enhance splicing. First, we obtained the recombinant virus which lost almost 70% of C-terminus by substituting Phe at residue 91 into termination codon (F91 stop). HMV-II cells infected with the recombinant virus containing F91 stop mutation showed that the ratio of M1 derived from spliced M mRNA / CM2 from unspliced M mRNA was reduced by 20%, a finding which suggests that the functional domain may be present in C-terminal region. The functional domain for enhancing splicing remains to be elucidated by generation NS1 mutants containing the various deletions in C-terminus.

我们最近使用表达PB 2、PB 1、P3、HE、NP、M和NS病毒RNA的7个Pol I质粒和表达PB 2、PB 1、P3、HE、NP、M1、CM 2、NS 1和NS的9个质粒建立了丙型流感病毒的反向遗传学，并成功地产生了重组丙型流感病毒。为了确定丙型流感病毒NS基因产物是否在前体mRNA剪接中发挥作用，我们尝试获得病毒，其中Leu 20被替换为终止密码子以拯救NS 1和NS 2表达很少的病毒。然而，病毒没有被拯救，这表明NS基因产物，NS 1和/或NS 2，是病毒复制所必需的。在瞬时表达系统中，M基因cDNA与NS 1蛋白共表达，M前mRNA的剪接率提高了30%，提示NS 1可能具有增强剪接的活性。然后，我们研究了NS 1特异性的C-末端区域（63-246个残基）是否具有增强剪接的功能结构域。首先，我们通过将第91位残基的Phe替换为终止密码子（F91终止），获得了C-末端丢失近70%的重组病毒。用含有F91终止突变的重组病毒感染HMV-Ⅱ细胞，发现剪接的M mRNA衍生的M1/未剪接的M mRNA衍生的CM 2的比值降低了20%，这一发现表明功能结构域可能存在于C端区域。增强剪接的功能结构域仍有待于通过产生在C-末端含有各种缺失的NS 1突变体来阐明。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A nationwide epidemic of influenza C virus infection in Japan in 2004

DOI：
10.1128/jcm.01555-06
发表时间：
2007-03-01
期刊：
JOURNAL OF CLINICAL MICROBIOLOGY
影响因子：
9.4
作者：
Matsuzaki, Yoko;Abiko, Chieko;Nishimura, Hidekazu
通讯作者：
Nishimura, Hidekazu

Conformational maturation of the nucleoprotein synthesized in influenza C virus-infected cells.

丙型流感病毒感染细胞中合成的核蛋白的构象成熟。

DOI：
发表时间：
2006
期刊：
Virus Res. 122
影响因子：
0
作者：
Umene K;et al.;Sugawara K
通讯作者：
Sugawara K

An outbreak of measles virus infection due to a genotype D9 at a junior high school in Yamagata, Japan in 2004.

2004 年，日本山形市一所初中爆发了 D9 基因型麻疹病毒感染。

DOI：
发表时间：
2005
期刊：
Jpn. J. Infec. Dis. 58
影响因子：
0
作者：
Takamasa Ueno;Masafumi Takiguchi;梅根健一;Matsumoto K;Tanaka T.;上野貴将;Nakashima K;Mizuta K
通讯作者：
Mizuta K

Isolation of an influenza C virus introduced into Japan by a traveler from Malaysia