Comprehensive analysis of ebola virus GP1 in viral entry.
Ebola virus infection is initiated by interactions between the viral glycoprotein GP1 and its cognate receptor(s), but little is known about the structure and function of GP1 in viral entry, partly due to the concern about safety when working with the live Ebola virus and the difficulty of manipulating the RNA genome of Ebola virus. In this study, we have used a human immunodeficiency virus-based pseudotyped virus as a surrogate system to dissect the role of Ebola virus GP1 in viral entry. Analysis of more than 100 deletion and amino acid substitution mutants of GP1 with respect to protein expression, processing, viral incorporation, and viral entry has allowed us to map the region of GP1 responsible for viral entry to the N-terminal 150 residues. Furthermore, six amino acids in this region have been identified as critical residues for early events in Ebola virus entry, and among these, three are clustered and are implicated as part of a potential receptor-binding pocket. In addition, substitutions of some 30 residues in GP1 are shown to adversely affect GP1 expression, processing, and viral incorporation, suggesting that these residues are involved in the proper folding and/or overall conformation of GP. Sequence comparison of the GP1 proteins suggests that the majority of the critical residues for GP folding and viral entry identified in Ebola virus GP1 are conserved in Marburg virus. These results provide information for elucidating the structural and functional roles of the filoviral glycoproteins and for developing potential therapeutics to block viral entry.
Manicassamy
Balaji
B
Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicago, E829 MSB, 835 S. Wolcott Ave., Chicago, IL 60612, USA.
Wang
Jizhen
J
Jiang
Haiqing
H
Rong
Lijun
L
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002460:Cell Line; D016384:Consensus Sequence; D029043:Ebolavirus / Q000502:physiology*; D017353:Gene Deletion; D006801:Humans; D007668:Kidney; D029024:Marburgvirus / Q000502:physiology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017510:Protein Folding; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D014759:Viral Envelope Proteins; C113189:envelope glycoprotein, Ebola virus