The Role of the Charged Residues of the GP2 Helical Regions in Ebola Entry

Haiqing Jiang; Jizhen Wang; Balaji Manicassamy; Santhakumar Manicassamy; Michael Caffrey; Lijun Rong

doi:10.1007/s12250-009-3015-6

Back

The Role of the Charged Residues of the GP2 Helical Regions in Ebola Entry

Journal article

Open access

Peer reviewed

The Role of the Charged Residues of the GP2 Helical Regions in Ebola Entry

Haiqing Jiang, Jizhen Wang, Balaji Manicassamy, Santhakumar Manicassamy, Michael Caffrey and Lijun Rong

Virologica Sinica, Vol.24(2), pp.121-135

04/2009

DOI: 10.1007/s12250-009-3015-6

PMCID: PMC3516429

PMID: 23227032

Files and links (1)

url

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3516429View

Published (Version of record) Open Access

Abstract

The glycoprotein (GP) of Ebola is the sole structural protein that forms the spikes on the viral envelope. The GP contains two subunits, GP1 and GP2, linked by a disulfide bond, which are responsible for receptor binding and membrane fusion, respectively. In this study, the full length of GP gene of Ebola Zaire species, 2028 base pairs in length, was synthesized using 38 overlapping oligonucleotides by multiple rounds of polymerase chain reaction (PCR). The synthesized GP gene was shown to be efficiently expressed in mammalian cells. Furthermore, an efficient HIV-based pseudotyping system was developed using the synthetic GP gene, providing a safe approach to dissecting the entry mechanism of Ebola viruses. Using this pseudotyping system and mutational analysis, the role of the charged residues in the GP2 helical regions was examined. It was found that substitutions of the most charged residues in the regions did not adversely affect GP expression, processing, or viral incorporation, however, most of the mutations greatly impaired the ability of GP to mediate efficient viral infection. These results demonstrate that these charged residues of GP2 play an important role in GP-mediated Ebola entry into its host cells. We propose that these charged residues are involved in forming the intermediate conformation(s) of GP in membrane fusion and Ebola entry.

Ebola Virus

GP2

Viral entry

Glycoprotein GP1

Charged residues

Details

Title: Subtitle: The Role of the Charged Residues of the GP2 Helical Regions in Ebola Entry
Creators: Haiqing Jiang - Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicag
Jizhen Wang - Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicag
Balaji Manicassamy - Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicag
Santhakumar Manicassamy - Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicag
Michael Caffrey - Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago College of Medicine, Chicago, IL 60612, USA
Lijun Rong - Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicag
Resource Type: Journal article
Publication Details: Virologica Sinica, Vol.24(2), pp.121-135
DOI: 10.1007/s12250-009-3015-6
PMID: 23227032
PMCID: PMC3516429
NLM abbreviation: Virol Sin
ISSN: 1674-0769
eISSN: 1995-820X
Grant note: R01 AI059570-04 || AI / National Institute of Allergy and Infectious Diseases Extramural Activities : NIAID
Language: English
Date published: 04/2009
Academic Unit: Microbiology and Immunology
Record Identifier: 9984083809002771

Metrics

18 Record Views