Arginine methylation regulates the function of a key herpesvirus nucleocytoplasmic shuttle protein

精氨酸甲基化调节关键疱疹病毒核细胞质穿梭蛋白的功能

• 批准号: BB/F012101/1
• 负责人: Adrian Whitehouse
• 金额: $ 39.77万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FF012101%2F1
• 关键词: Arginine; methylation; regulates; function; key

mRNA is transcribed in the nucleus and must be exported from the nucleus into the cytoplasm to get translated into a protein. In the nucleus, the RNA has to undergo a series of processing events prior to export and these are performed by a number of multi-protein complexes. However, how these complexes are recruited to the RNA and how each multi-protein complex is regulated to perform its specific function at the correct time is unknown. Herein, we aim to investigate how the function of one of these RNA binding multi-protein complexes is regulated using a viral model. We have previously demonstrated that the herpesvirus ORF57 protein is important for exporting herpesvirus mRNA out of the nucleus to get translated. ORF57 functions by binding to the herpesvirus RNA and specifically recruiting cellular proteins onto the viral RNA, which are essential for its correct processing and export. This system has uniquely identified that a multi-protein complex called hTREX is the only cellular protein complex which is required by ORF57 for herpesvirus mRNA nuclear export. Moreover, we have demonstrated that once hTREX is bound by ORF57 to the herpesvirus mRNA, the whole complex is transported through a domain within the nucleus called the nucleolus, before it is exported from the nucleus into the cytoplasm. Therefore, ORF57 is a multifunctional protein which sequentially, binds the viral RNA, then recruits hTREX to the viral RNA, then transports this complex to the nucleolus, then allows the complex to exit the nucleus. But what tells ORF57 to perform its many functions in the correct manner is unknown. This functional diversity of a protein can be controlled by different chemical modifications to the protein, such as adding or taking away a phosphate, acetate or methyl group on the protein. Interestingly, we have demonstrated that the herpesvirus ORF57 protein undergoes one such modification. We have shown that ORF57 can be methylated at arginine residues. Therefore, we aim to determine if adding or taking away methyl groups affects ORF57's function and in turn regulates the multi-protein complex responsible for exporting viral mRNA from the nucleus. To assess the role of methylation we will first identify which residues of ORF57 are methylated. We will then mutate these, so that ORF57 cannnot undergo methylation. This will then allow us to assess what effect it has upon the functioning of the ORF57 protein. Secondly, we will identify which cellular enzymes are responsible for performing this post-translational modification on the viral ORF57 protein. Finally, we have preliminary data to suggest that taking away the methyl group on ORF57 (or demethylating ORF57), is the trigger to tell ORF57 to exit the nucleus. We believe this happens in the nucleolus and we will identify which enzyme in the nucleolus performs this function. These data will provide a better understanding of how multi-protein complexes function to enable RNAs to exit the nucleus and also provide a better understanding of how viruses function in order to identify new antiviral targets in the longer term.

mRNA is transcribed in the nucleus and must be exported from the nucleus into the cytoplasm to get translated into a protein. In the nucleus, the RNA has to undergo a series of processing events prior to export and these are performed by a number of multi-protein complexes. However, how these complexes are recruited to the RNA and how each multi-protein complex is regulated to perform its specific function at the correct time is unknown. Herein, we aim to investigate how the function of one of these RNA binding multi-protein complexes is regulated using a viral model. We have previously demonstrated that the herpesvirus ORF57 protein is important for exporting herpesvirus mRNA out of the nucleus to get translated. ORF57 functions by binding to the herpesvirus RNA and specifically recruiting cellular proteins onto the viral RNA, which are essential for its correct processing and export. This system has uniquely identified that a multi-protein complex called hTREX is the only cellular protein complex which is required by ORF57 for herpesvirus mRNA nuclear export. Moreover, we have demonstrated that once hTREX is bound by ORF57 to the herpesvirus mRNA, the whole complex is transported through a domain within the nucleus called the nucleolus, before it is exported from the nucleus into the cytoplasm. Therefore, ORF57 is a multifunctional protein which sequentially, binds the viral RNA, then recruits hTREX to the viral RNA, then transports this complex to the nucleolus, then allows the complex to exit the nucleus. But what tells ORF57 to perform its many functions in the correct manner is unknown. This functional diversity of a protein can be controlled by different chemical modifications to the protein, such as adding or taking away a phosphate, acetate or methyl group on the protein. Interestingly, we have demonstrated that the herpesvirus ORF57 protein undergoes one such modification. We have shown that ORF57 can be methylated at arginine residues. Therefore, we aim to determine if adding or taking away methyl groups affects ORF57's function and in turn regulates the multi-protein complex responsible for exporting viral mRNA from the nucleus. To assess the role of methylation we will first identify which residues of ORF57 are methylated. We will then mutate these, so that ORF57 cannnot undergo methylation. This will then allow us to assess what effect it has upon the functioning of the ORF57 protein. Secondly, we will identify which cellular enzymes are responsible for performing this post-translational modification on the viral ORF57 protein. Finally, we have preliminary data to suggest that taking away the methyl group on ORF57 (or demethylating ORF57), is the trigger to tell ORF57 to exit the nucleus. We believe this happens in the nucleolus and we will identify which enzyme in the nucleolus performs this function. These data will provide a better understanding of how multi-protein complexes function to enable RNAs to exit the nucleus and also provide a better understanding of how viruses function in order to identify new antiviral targets in the longer term.

项目成果

Recruitment of the complete hTREX complex is required for Kaposi's sarcoma-associated herpesvirus intronless mRNA nuclear export and virus replication.

• DOI: 10.1371/journal.ppat.1000194
• 发表时间: 2008-10
• 期刊: PLOS PATHOGENS
• 影响因子: 6.7
• 作者: Boyne, James R.;Colgan, Kevin J.;Whitehouse, Adrian
• 通讯作者: Whitehouse, Adrian

Mutation of a C-terminal motif affects Kaposi's sarcoma-associated herpesvirus ORF57 RNA binding, nuclear trafficking, and multimerization.

C 末端基序的突变会影响卡波西肉瘤相关疱疹病毒 ORF57 RNA 结合、核运输和多聚化。

• DOI: 10.1128/jvi.00138-11
• 发表时间: 2011
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Taylor A

An interaction between KSHV ORF57 and UIF provides mRNA-adaptor redundancy in herpesvirus intronless mRNA export.

• DOI: 10.1371/journal.ppat.1002138
• 发表时间: 2011-07
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Jackson BR;Boyne JR;Noerenberg M;Taylor A;Hautbergue GM;Walsh MJ;Wheat R;Blackbourn DJ;Wilson SA;Whitehouse A
• 通讯作者: Whitehouse A

Kaposi's sarcoma-associated herpesvirus ORF57 protein interacts with PYM to enhance translation of viral intronless mRNAs.

• DOI: 10.1038/emboj.2010.77
• 发表时间: 2010-06-02
• 期刊: The EMBO journal
• 作者: Boyne JR;Jackson BR;Taylor A;Macnab SA;Whitehouse A
• 通讯作者: Whitehouse A