The paramyxovirus polymerase complex as a target for next-generation anti-paramyxovirus therapeutics

Cox, Robert; Plemper, Richard K.

doi:10.3389/fmicb.2015.00459

Cited by 47 publications

(37 citation statements)

References 155 publications

(240 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…According to current models of replication/ transcription of negative strand non-segmented RNA viruses, the template RNA is covered by N protein (NP) and only 2-3 NP molecules are assumed to be displaced simultaneously at the site of transcription, giving access to not more than 30 nucleotides. 18,19 Yet, at present it cannot be excluded that larger regions of RNA become transiently accessible during EBOV transcriptional initiation (e.g. by VP30 displacing NP), also taking into account that the leader promoter is bipartite in EBOV, and thus more extended, than in vesicular stomatitis virus (VSV) or respiratory syncytial virus (RSV); furthermore, Ebola VP30 is an exceptionally large and unique viral transcription factor within the Mononegavirales, which leaves the possibility open that temporary RNA accessibilities are different in the EBOV system.…”

Section: Discussionmentioning

confidence: 99%

RNA binding specificity of Ebola virus transcription factor VP30

et al. 2016

View full text Add to dashboard Cite

The transcription factor VP30 of the non-segmented RNA negative strand Ebola virus balances viral transcription and replication. Here, we comprehensively studied RNA binding by VP30. Using a novel VP30:RNA electrophoretic mobility shift assay, we tested truncated variants of 2 potential natural RNA substrates of VP30 -the genomic Ebola viral 3 0 -leader region and its complementary antigenomic counterpart (each »155 nt in length) -and a series of other non-viral RNAs. Based on oligonucleotide interference, the major VP30 binding region on the genomic 3 0 -leader substrate was assigned to the internal expanded single-stranded region (» nt 125-80). Best binding to VP30 was obtained with ssRNAs of optimally » 40 nt and mixed base composition; underrepresentation of purines or pyrimidines was tolerated, but homopolymeric sequences impaired binding. A stem-loop structure, particularly at the 3 0 -end or positioned internally, supports stable binding to VP30. In contrast, dsRNA or RNAs exposing large internal loops flanked by entirely helical arms on both sides are not bound. Introduction of a 5-Cap(0) structure impaired VP30 binding. Also, ssDNAs bind substantially weaker than isosequential ssRNAs and heparin competes with RNA for binding to VP30, indicating that ribose 2 0 -hydroxyls and electrostatic contacts of the phosphate groups contribute to the formation of VP30:RNA complexes. Our results indicate a rather relaxed RNA binding specificity of filoviral VP30, which largely differs from that of the functionally related transcription factor of the Paramyxoviridae which binds to ssRNAs as short as 13 nt with a preference for oligo(A) sequences.

show abstract

Section: Discussionmentioning

confidence: 99%

RNA binding specificity of Ebola virus transcription factor VP30

et al. 2016

View full text Add to dashboard Cite

show abstract

“…The hRSV RNA-dependent RNA complex (RdRp) constitutes a virus-specific target with specific protein-protein interactions that have not all been investigated in detail (7). It uses the nonsegmented single-stranded negative sense RNA genome of hRSV as a template.…”

mentioning

confidence: 99%

New Insights into Structural Disorder in Human Respiratory Syncytial Virus Phosphoprotein and Implications for Binding of Protein Partners

Pereira¹,

Cardone²,

Lassoued³

et al. 2017

Journal of Biological Chemistry

116

View full text Add to dashboard Cite

Edited by Charles E. SamuelPhosphoprotein is the main cofactor of the viral RNA polymerase of Mononegavirales. It is involved in multiple interactions that are essential for the polymerase function. Most prominently it positions the polymerase complex onto the nucleocapsid, but also acts as a chaperone for the nucleoprotein. Mononegavirales phosphoproteins lack sequence conservation, but contain all large disordered regions. We show here that Nand C-terminal intrinsically disordered regions account for 80% of the phosphoprotein of the respiratory syncytial virus. But these regions display marked dynamic heterogeneity. Whereas almost stable helices are formed C terminally to the oligomerization domain, extremely transient helices are present in the N-terminal region. They all mediate internal long-range contacts in this non-globular protein. Transient secondary elements together with fully disordered regions also provide protein binding sites recognized by the respiratory syncytial virus nucleoprotein and compatible with weak interactions required for the processivity of the polymerase. Human respiratory syncytial virus (hRSV),3 a member of the family Pneumoviridae (1) and order Mononegavirales (MNV), is the main viral cause of lower respiratory tract illness worldwide, and the main agent responsible for bronchiolitis and pneumonia in infants (2). All children have been infected by the age of two, requiring hospitalization in ϳ5% cases (3). Elderly and immunocompromised adults are also at increased risk. No efficient treatment is presently available for hRSV (4), and vaccination is challenging due to complex immunogenicity (5). The search for hRSV antiviral drugs directed toward specific viral functions is therefore still ongoing (6).The hRSV RNA-dependent RNA complex (RdRp) constitutes a virus-specific target with specific protein-protein interactions that have not all been investigated in detail (7). It uses the nonsegmented single-stranded negative sense RNA genome of hRSV as a template. In infected cells, the viral RdRp is found in specific inclusion bodies (8), which have been shown to be transcription and replication centers for other Mononegavirales, e.g. rabies (9) and vesicular stomatitis viruses (10). The apo RdRp complex is composed a minima of the large catalytic subunit (L) and its essential cofactor, the phosphoprotein (P) (11, 12). The P protein plays a central role in the RdRp by interacting with all main RdRp components. During transcription and replication it tethers the L protein to the nucleocapsid (NC), consisting of the genomic RNA packaged by the nucleoprotein (N), by direct interaction with N (13-16). hRSV P also binds to the transcription antitermination factor M2-1 (17-19). Phosphorylation of P has been proposed to regulate these interactions, although it is not essential for replication (20 -22). P also acts as a chaperone for neo-synthesized N by forming an N 0 ⅐P complex that preserves N in a monomeric and RNA-free state (23). We have shown previously that formation of hRSV NC⅐P and ...

show abstract

“…Following cell entry, the genomic RNPs are released into the cytosol and the encapsidated viral RNA serves as a template of the RdRP complex for both transcription and replication [5]. Transcription begins at the 3′ end of the genome and viral genes are transcribed in the 3′ to 5′direction with a sequential “stop–start” mechanism.…”

Section: Genome and Lifecyclementioning

confidence: 99%

“…The virion of MeV is a pleomorphic or spherical particle with a diameter ranging from 120 to 1000 nm and has two major structural components: one is the helical ribonucleoprotein (RNP) core formed by the association of the nucleoprotein (N), phosphoprotein (P) and large protein (L) with the viral genome, the other is the cellular membrane-derived lipid envelope surrounding the RNP core [1,4,5]. The active RNP complex is responsible for initiating primary transcription after cell entry as well as counteracting the host interferon (IFN) signaling pathway [5,6,7].…”

Section: Introductionmentioning

confidence: 99%

“…The active RNP complex is responsible for initiating primary transcription after cell entry as well as counteracting the host interferon (IFN) signaling pathway [5,6,7]. The MeV RNP is bound by the matrix protein (M), and then covered by the lipid envelope containing two spike glycoproteins, F and H proteins, which are primarily responsible for membrane fusion and receptor attachment, respectively [8].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Host–Pathogen Interactions in Measles Virus Replication and Anti-Viral Immunity

Jiang

Qin

Chen

2016

Viruses

View full text Add to dashboard Cite

The measles virus (MeV) is a contagious pathogenic RNA virus of the family Paramyxoviridae, genus Morbillivirus, that can cause serious symptoms and even fetal complications. Here, we summarize current molecular advances in MeV research, and emphasize the connection between host cells and MeV replication. Although measles has reemerged recently, the potential for its eradication is promising with significant progress in our understanding of the molecular mechanisms of its replication and host-pathogen interactions.

show abstract

The paramyxovirus polymerase complex as a target for next-generation anti-paramyxovirus therapeutics

Cited by 47 publications

References 155 publications

RNA binding specificity of Ebola virus transcription factor VP30

RNA binding specificity of Ebola virus transcription factor VP30

New Insights into Structural Disorder in Human Respiratory Syncytial Virus Phosphoprotein and Implications for Binding of Protein Partners

Host–Pathogen Interactions in Measles Virus Replication and Anti-Viral Immunity

Contact Info

Product

Resources

About