title: Deletion of the D domain of the human parainfluenza virus type 3 (HPIV3) PD protein results in decreased viral RNA synthesis and beta interferon (IFN-β) expression.
abstract: The human parainfluenza virus type 3 (HPIV3) phosphoprotein (P) gene is unusual as it contains an editing site where nontemplated ribonucleotide residues can be inserted. This RNA editing can lead to the expression of the viral P, PD, putative W, and theoretical V protein from a single gene. Although the HPIV3 PD protein has been detected, its function and those of the W and V proteins are poorly understood. Therefore, we first used reverse genetics techniques to construct and rescue a recombinant (r)HPIV3 clone with a polyhistidine sequence at the 5' end of the P gene for tagged protein detection. Western blot analysis demonstrated the presence of the P, PD, and W proteins, but no V protein was detected. Then, we functionally studied the D domain of the PD protein by constructing two rHPIV3 knockout clones that are deficient in the expression of the D domain. Results from growth kinetic studies with infected MA-104 and A596 cells showed that viral replication of the two knockout viruses (rHPIV3-ΔES and rHPIV3-ΔD) was comparable to that of the parental virus in both cell lines. However, viral mRNA transcription and genomic replication was significantly reduced. Furthermore, cytokine/chemokine profiles of A549 cells infected with either knockout virus were unchanged or showed lower levels compared to those from cells infected with the parental virus. These data suggest that the D domain of the PD protein may play a luxury role in HPIV3 RNA synthesis and may also be involved in disrupting the expression of beta interferon.
authors.affiliation: Department of Animal, Dairy, and Veterinary Sciences, Institute for Antiviral Research, Utah State University, 5600 Old Main Hill, Logan, UT 84322-5600, USA.
authors.forename: Jason P
authors.initials: JP
authors.lastname: Roth
authors.forename: Joseph K-K
authors.initials: JK
authors.lastname: Li
authors.forename: John D
authors.initials: JD
authors.lastname: Morrey
authors.forename: Dale L
authors.initials: DL
authors.lastname: Barnard
authors.forename: Almut H
authors.initials: AH
authors.lastname: Vollmer
mesh_terms: D002460:Cell Line; D015536:Down-Regulation; D006801:Humans; D016899:Interferon-beta / Q000235:genetics* / Q000276:immunology; D010224:Parainfluenza Virus 3, Human / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D010253:Respirovirus Infections / Q000235:genetics* / Q000276:immunology / Q000821:virology; D017384:Sequence Deletion; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: C095185:P protein, Human parainfluenza virus 3; D010750:Phosphoproteins; D012367:RNA, Viral; D014764:Viral Proteins; D016899:Interferon-beta