Interaction Interface of Mason-Pfizer Monkey Virus Matrix and Envelope Proteins

0533120 - ÚOCHB 2021 RIV US eng J - Journal Article
Prchal, J. - Sýs, Jakub - Junková, Petra - Lipov, J. - Ruml, T.
Interaction Interface of Mason-Pfizer Monkey Virus Matrix and Envelope Proteins.
Journal of Virology. Roč. 94, č. 20 (2020), č. článku e01146-20. ISSN 0022-538X. E-ISSN 1098-5514
R&D Projects: GA MŠMT(CZ) EF16_019/0000729
Institutional support: RVO:61388963
Keywords : Env * M-PMV * retrovirus * cytoplasmic tail * protein interactions
OECD category: Analytical chemistry
Impact factor: 5.103, year: 2020
Method of publishing: Limited access
https://doi.org/10.1128/jvi.01146-20

Retroviral envelope glycoprotein (Env) is essential for the specific recognition of the host cell and the initial phase of infection. As reported for human immunodeficiency virus (HIV), the recruitment of Env into a retroviral membrane envelope is mediated through its interaction with a Gag polyprotein precursor of structural proteins. This interaction, occurring between the matrix domain (MA) of Gag and the cytoplasmic tail (CT) of the transmembrane domain of Env, takes place at the host cell plasma membrane. To determine whether the MA of Mason-Pfizer monkey virus (M-PMV) also interacts directly with the CT of Env, we mimicked the in vivo conditions in an in vitro experiment by using a CT in its physiological trimeric conformation mediated by the trimerization motif of the GCN4 yeast transcription factor. The MA protein was used at the concentration shifting the equilibrium to its trimeric form. The direct interaction between MA and CT was confirmed by a pulldown assay. Through the combination of nuclear magnetic resonance (NMR) spectroscopy and protein cross-linking followed by mass spectrometry analysis, the residues involved in mutual interactions were determined. NMR has shown that the C terminus of the CT is bound to the C-terminal part of MA. In addition, protein cross-linking confirmed the close proximity of the N-terminal part of CT and the N terminus of MA, which is enabled in vivo by their location at the membrane. These results are in agreement with the previously determined orientation of MA on the membrane and support the already observed mechanisms of M-PMV virus-like particle transport and budding.
Permanent Link: http://hdl.handle.net/11104/0311741