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Identification of a Membrane Binding Peptide in the Envelope Protein of MHV Coronavirus
Entedar A J Alsaadi1,2, Benjamin W Neuman3, Ian M Jones1
1School of Biological Sciences, University of Reading, Reading RG6 6AJ, UK.
Abstract:
Coronaviruses (CoVs) are enveloped, positive sense, single strand RNA viruses that cause respiratory, intestinal and neurological diseases in mammals and birds. Following replication, CoVs assemble on intracellular membranes including the endoplasmic reticulum Golgi intermediate compartment (ERGIC) where the envelope protein (E) functions in virus assembly and release. In consequence, E potentially contains membrane-modifying peptides. To search for such peptides, the E coding sequence of Mouse Hepatitis Virus (MHV) was inspected for its amino acid conservation, proximity to the membrane and/or predicted amphipathic helices. Peptides identified in silico were synthesized and tested for membrane-modifying activity in the presence of giant unilamellar vesicles (GUVs) consisting of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), sphingomyelin and cholesterol. To confirm the presence of membrane binding peptides identified in the context of a full-length E protein, the wild type and a number of mutants in the putative membrane binding peptide were expressed in Lenti-X-293T mammalian and insect cells, and the distribution of E antigen within the expressing cell was assessed. Our data identify a role for the post-transmembrane region of MHV E in membrane binding.
Insights
Coronaviruses assemble on cell membranes, aided by the envelope protein (E). Researchers found that a specific region of the Mouse Hepatitis Virus E protein binds to membranes, impacting virus assembly.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Coronaviruses (CoVs) are RNA viruses causing various diseases in mammals and birds.
- CoVs assemble within cells, utilizing intracellular membranes like the ERGIC.
- The viral envelope protein (E) is crucial for CoV assembly and release.
Purpose of the Study:
- To identify potential membrane-modifying peptides within the Mouse Hepatitis Virus (MHV) E protein.
- To investigate the role of these peptides in virus assembly and membrane interaction.
Main Methods:
- In silico analysis of the MHV E protein sequence for conserved, membrane-proximal, amphipathic helical regions.
- Synthesis and in vitro testing of identified peptides for membrane-modifying activity using GUVs.
- Expression of wild-type and mutant MHV E proteins in mammalian and insect cells to assess cellular localization.
Main Results:
- Identification of potential membrane-binding peptides within the MHV E protein sequence.
- Experimental confirmation of membrane binding activity for synthesized peptides.
- Demonstration that the post-transmembrane region of MHV E is involved in membrane binding.
Conclusions:
- The post-transmembrane region of the MHV E protein plays a significant role in membrane binding.
- This finding contributes to understanding the molecular mechanisms of coronavirus assembly and release.
