It is the objective of the work proposed in this grant application to enhance our understanding of the mechanisms by which poliovirus (PV), an enterovirus belonging to the Picornaviridae, encapsidates its genome. This virus family includes a large number of human and animal pathogens that cause more than 6 billion human infections worldwide each year. These infections lead to a variety of diseases ranging from the mild (common cold) to the serious (poliomyelitis). In spite of research for many years the details of most steps in the life cycle of PV remain unknown. However, the proliferation of PV remains an important medical issue because epidemic PV infections are expected to occur even after the circulation of wt PV is interrupted globally. This proposal can be divided into 3 parts.
The first aim deals with the development of new genetic tools to study the genetics of encapsidation of chimeras constructed from PV and the closely related C-cluster coxsackie A viruses. We expect that the morphogenesis phenotypes of the chimeric viruses will be very useful in analyzing the interaction of capsid and nonstructural proteins during encapsidation.
The second aim of the proposal deals with two specific inhibitors of enterovirus encapsidation, hydantoin and L-buthionine-sulfoximine (BSO), whose effect is expected to block encapsidation at different stages. In this study PV, CAVs and PV/C-CAV chimeras will be used to identify and analyze escape mutants in proteins involved in encapsidation. In our third aim we propose to analyze the role in morphogenesis of non-structural proteins 2CATPase, 3CDpro, VPg, and, and search for an elusive RNA encapsidation signal. We plan to use both genetic and biochemical studies to analyze the role of nonstructural proteins 2CATPase and 3CDpro in encapsidation. The role of VPg will be tested by a large-scale scan of hundreds of VPg mutants with the aim of finding replication positive but encapsidation negative mutants. Finally, we will use a novel strategy (codon-pair optimization) to scan the PV RNA for an elusive encapsidation signal. Since encapsidation is a uniquely viral process an understanding of its mechanism will aid the development of antiviral drugs that target this particular step in the enteroviral life cycle. The 2CATPase and 3CDpro proteins are highly conserved among enteroviruses hence they provide an excellent target for drug development to treat multiple enteroviral diseases. It is believed that these studies and results will be of interest not only to those investigators who study the encapsidation of other enteroviruses but also to those who are interested in picornaviruses or RNA viruses in general.
Picornaviridae cause about 6 billion human infections a year worldwide, some leading to serious diseases. Poliovirus, a member of the enterovirus genus, is the prototype of this virus family. The project proposed in this application deals with the mechanism of encapsidation of enteroviruses, particularly of poliovirus and of the closely related c-cluster enteroviruses. Our understanding of the mechanism of encapsidation is expected to facilitate the development of new drugs to treat enteroviral infections.
|Chen, Ying-Han; Du, WenLi; Hagemeijer, Marne C et al. (2015) Phosphatidylserine vesicles enable efficient en bloc transmission of enteroviruses. Cell 160:619-630|
|Paul, Aniko V; Wimmer, Eckard (2015) Initiation of protein-primed picornavirus RNA synthesis. Virus Res 206:12-26|
|Jiang, Ping; Liu, Ying; Ma, Hsin-Chieh et al. (2014) Picornavirus morphogenesis. Microbiol Mol Biol Rev 78:418-37|
|Ma, Hsin-Chieh; Liu, Ying; Wang, Chunling et al. (2014) An interaction between glutathione and the capsid is required for the morphogenesis of C-cluster enteroviruses. PLoS Pathog 10:e1004052|
|Schein, Catherine H; Bowen, David M; Lewis, Jessica A et al. (2012) Physicochemical property consensus sequences for functional analysis, design of multivalent antigens and targeted antivirals. BMC Bioinformatics 13 Suppl 13:S9|
|Wang, Chunling; Jiang, Ping; Sand, Claire et al. (2012) Alanine scanning of poliovirus 2CATPase reveals new genetic evidence that capsid protein/2CATPase interactions are essential for morphogenesis. J Virol 86:9964-75|
|Song, Yutong; Liu, Ying; Ward, Charles B et al. (2012) Identification of two functionally redundant RNA elements in the coding sequence of poliovirus using computer-generated design. Proc Natl Acad Sci U S A 109:14301-7|
|Toyoda, Hidemi; Wimmer, Eckard; Cello, Jeronimo (2011) Oncolytic poliovirus therapy and immunization with poliovirus-infected cell lysate induces potent antitumor immunity against neuroblastoma in vivo. Int J Oncol 38:81-7|
|Wimmer, Eckard; Paul, Aniko V (2011) Synthetic poliovirus and other designer viruses: what have we learned from them? Annu Rev Microbiol 65:583-609|
|Liu, Ying; Wang, Chunling; Mueller, Steffen et al. (2010) Direct interaction between two viral proteins, the nonstructural protein 2C and the capsid protein VP3, is required for enterovirus morphogenesis. PLoS Pathog 6:e1001066|
Showing the most recent 10 out of 134 publications