Article ; Online: Specific Residues in the C-Terminal Domain of the Human Metapneumovirus Phosphoprotein Are Indispensable for Formation of Viral Replication Centers and Regulation of the Function of the Viral Polymerase Complex.
2023 Volume 97, Issue 5, Page(s) e0003023
Abstract: Human metapneumovirus (HMPV) is a negative-strand RNA virus that frequently causes respiratory tract infections in infants, the elderly, and the immunocompromised. A hallmark of HMPV infection is the formation of membraneless, liquid-like replication and ...
| Abstract | Human metapneumovirus (HMPV) is a negative-strand RNA virus that frequently causes respiratory tract infections in infants, the elderly, and the immunocompromised. A hallmark of HMPV infection is the formation of membraneless, liquid-like replication and transcription centers in the cytosol termed inclusion bodies (IBs). The HMPV phosphoprotein (P) and nucleoprotein (N) are the minimal viral proteins necessary to form IB-like structures, and both proteins are required for the viral polymerase to synthesize RNA during infection. HMPV P is a homotetramer with regions of intrinsic disorder and has several known and predicted phosphorylation sites of unknown function. In this study, we found that the P C-terminal intrinsically disordered domain (CTD) must be present to facilitate IB formation with HMPV N, while either the N-terminal intrinsically disordered domain or the central oligomerization domain was dispensable. Alanine substitution at a single tyrosine residue within the CTD abrogated IB formation and reduced coimmunoprecipitation with HMPV N. Mutations to C-terminal phosphorylation sites revealed a potential role for phosphorylation in regulating RNA synthesis and P binding partners within IBs. Phosphorylation mutations which reduced RNA synthesis in a reporter assay produced comparable results in a recombinant viral rescue system, measured as an inability to produce infectious viral particles with genomes containing these single P mutations. This work highlights the critical role HMPV P plays in facilitating a key step of the viral life cycle and reveals the potential role for phosphorylation in regulating the function of this significant viral protein. |
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| MeSH term(s) | Aged ; Humans ; Cell Line ; Metapneumovirus/physiology ; Nucleotidyltransferases ; Paramyxoviridae Infections/virology ; Phosphoproteins/genetics ; Phosphoproteins/metabolism ; Respiratory Tract Infections ; RNA ; Viral Proteins/genetics ; Viral Proteins/metabolism ; Viral Replication Compartments/metabolism ; Virus Replication ; Inclusion Bodies, Viral/metabolism |
| Chemical Substances | Nucleotidyltransferases (EC 2.7.7.-) ; Phosphoproteins ; RNA (63231-63-0) ; Viral Proteins |
| Language | English |
| Publishing date | 2023-04-24 |
| Publishing country | United States |
| Document type | Journal Article ; Research Support, N.I.H., Extramural |
| ZDB-ID | 80174-4 |
| ISSN | 1098-5514 ; 0022-538X |
| ISSN (online) | 1098-5514 |
| ISSN | 0022-538X |
| DOI | 10.1128/jvi.00030-23 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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