Reticulon 3 binds the 2C protein of enterovirus 71 and is required for viral replication.
Enterovirus 71 is an enterovirus of the family Picornaviridae. The 2C protein of poliovirus, a relative of enterovirus 71, is essential for viral replication. The poliovirus 2C protein is associated with host membrane vesicles, which form viral replication complexes where viral RNA synthesis takes place. We have now identified a host-encoded 2C binding protein called reticulon 3, which we found to be associated with the replication complex through direct interaction with the enterovirus 71-encoded 2C protein. We observed that the N terminus of the 2C protein, which has both RNA- and membrane-binding activity, interacted with reticulon 3. This region of interaction was mapped to its reticulon homology domain, whereas that of 2C was encoded by the 25th amino acid, isoleucine. Reticulon 3 could also interact with the 2C proteins encoded by other enteroviruses, such as poliovirus and coxsackievirus A16, implying that it is a common factor for such viral replication. Reduced production of reticulon 3 by RNA interference markedly reduced the synthesis of enterovirus 71-encoded viral proteins and replicative double-stranded RNA, reducing plaque formation and apoptosis. Furthermore, reintroduction of nondegradable reticulon 3 into these knockdown cells rescued enterovirus 71 infectivity, and viral protein and double-stranded RNA synthesis. Thus, reticulon 3 is an important component of enterovirus 71 replication, through its potential role in modulation of the sequential interactions between enterovirus 71 viral RNA and the replication complex.
Tang
Wen-Fang
WF
Department of Biochemistry and Chang Gung Bioinformatics Center, Chang Gung University, Kweishan, Taoyuan 333, Taiwan.
Yang
Shing-Ying
SY
Wu
Bin-Wen
BW
Jheng
Jia-Rong
JR
Chen
Yin-Li
YL
Shih
Chung-Hsuan
CH
Lin
Kwang-Huei
KH
Lai
Hsin-Chi
HC
Tang
Petrus
P
Horng
Jim-Tong
JT
D000818:Animals; D017209:Apoptosis; D002352:Carrier Proteins / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004770:Enterovirus / Q000502:physiology*; D017353:Gene Deletion; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000172:deficiency / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000172:deficiency / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012330:RNA, Double-Stranded / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D016601:RNA-Binding Proteins / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014779:Virus Replication / Q000502:physiology*
D002352:Carrier Proteins; D008565:Membrane Proteins; D046912:Multiprotein Complexes; D009419:Nerve Tissue Proteins; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D016601:RNA-Binding Proteins; C476198:RTN3 protein, human; D017361:Viral Nonstructural Proteins; C076023:2C protein, viral