Inhibition of measles virus minireplicon-encoded reporter gene expression by V protein

Witko, Susan E.; Kotash, Cheryl S.; Sidhu, M. S.; Udem, Stephen A.; Parks, Christopher L.

doi:10.1016/j.virol.2005.12.019

Cited by 33 publications

(27 citation statements)

References 58 publications

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“…We report here another role for this rubulavirus V protein, namely, as a negative regulator of genome replication. To date, the roles of the SeV, MeV, CDV, PIV5, and hPIV2 V proteins in viral RNA synthesis have been examined by using a similar transient-expression approach (32,62) or by examining defective interfering particle genome replication (7,20). In all cases, these V proteins were found to act as negative regulators.…”

Section: Discussionmentioning

confidence: 99%

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Human Parainfluenza Virus Type 2 V Protein Inhibits Genome Replication by Binding to the L Protein: Possible Role in Promoting Viral Fitness

Nishio

Ohtsuka

Tsurudome

et al. 2008

J Virol

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The human parainfluenza virus type 2 (hPIV2) V protein plays important roles in inhibiting the host interferon response and promoting virus growth, but its role in hPIV2 replication and transcription is not clear. A green fluorescent protein (GFP)-expressing a negative-sense minigenomic construct of hPIV2 has been established by standard technology, with helper plasmids expressing the nucleocapsid protein (NP), phosphoprotein (P), and large RNA polymerase (L) protein, to examine the role of V protein. We found that the simultaneous expression of wild-type V protein in the minigenome system inhibited GFP expression, at least in part, by inhibiting minigenome replication. In contrast, expression of C terminally truncated or mutant hPIV2 V proteins had no effect. Moreover, the V protein of simian virus 41, the rubulavirus most closely related virus to hPIV2, also inhibited GFP expression, whereas that of PIV5, a more distantly related rubulavirus, did not. Using these other rubulavirus V proteins, as well as various mutant hPIV2 V proteins, we found that the ability of V protein to inhibit GFP expression correlated with its ability to bind to L protein via its C-terminal V protein-specific region, but there was no correlation with NP binding. A possible role for this inhibition of genome replication in promoting viral fitness is discussed.Human parainfluenza virus type 2 (hPIV2) is a member of the Rubulavirus genus of the family Paramyxoviridae. This family includes many well-known human and animal pathogens, such as Sendai virus (SeV), hPIV types 1 to 4, simian virus 41 (SV41), parainfluenza virus type 5 (PIV5; formerly known as SV5), mumps virus, Newcastle disease virus, measles virus (MeV), and respiratory syncytial virus, as well as important emerging viruses such as Hendra and Nipah viruses. The negative-stranded RNA genome of hPIV2 is 15,654 nucleotides long and encodes seven viral proteins from six genes (30). The nucleocapsid protein (NP), phosphoprotein (P), and large RNA polymerase (L) protein are important for transcription and replication of the viral RNA genome. All viruses of the Paramyxoviridae (with the notable exception of hPIV1) contain an mRNA-editing site at which G residues are inserted into the P gene mRNA in a programmed manner during its synthesis. In respiroviruses and morbilliviruses, the P mRNA is a faithful copy of the genome RNA, and the V mRNA results from the insertion of one additional pseudotemplated G nucleotide. In only rubulaviruses, it is the V mRNA that is a faithful transcript of the V/P gene, whereas the P mRNA is synthesized through a cotranscriptional insertion of two pseudotemplated G residues. Thus, the N-terminal 164 amino acids (aa) of the V and P proteins are common, while their C termini are unique (43). Since insertion of the G residues in hPIV2 occurs ca. 50% of the time, roughly equal amounts of V and P mRNAs are produced. The C termini of the V proteins contain seven invariant cysteines that bind two atoms of zinc and is ca. 50% identical in sequence among all par...

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Section: Discussionmentioning

confidence: 99%

“…The RPV V and C proteins were found to interact with the L protein (54). Recently, the negative modulatory activity of V proteins encoded by PIV5 and MeV has been reproduced in transient minireplicon expression systems (32,62). However, the mechanisms of the V protein inhibition of these minigenome systems are not clear.…”

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confidence: 99%

Human Parainfluenza Virus Type 2 V Protein Inhibits Genome Replication by Binding to the L Protein: Possible Role in Promoting Viral Fitness

Nishio

Ohtsuka

Tsurudome

et al. 2008

J Virol

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“…Whilst the C proteins of NiV and of other paramyxoviruses are unique and share no sequence similarity with the P protein (Lamb & Kolakofsky, 2001), the V and W proteins share an amino-terminal 407 aa domain with P and each possesses a unique carboxyl-terminal domain consisting of 52 aa for V and 47 aa for W . The sequences of the P proteins of paramyxoviruses are not well conserved (Baron et al, 1993) and, whilst a V protein is not expressed by all paramyxoviruses, the unique carboxylterminal domain of V is more conserved than the aminoterminal domain shared with P (Galinski et al, 1992; Matsuoka et al, 1991;Witko et al, 2006). The C, V and W proteins of paramyxoviruses are known to play multiple roles in the viral life cycle, functioning as interferon antagonists (Didcock et al, 1999;Goodbourn et al, 2000;He et al, 2002;Komatsu et al, 2004;Lin et al, 2005 Shaffer et al, 2003), in addition to regulating viral transcription and replication both in vitro and in vivo (Bankamp et al, 2005;Baron & Barrett, 2000;Curran et al, 1991Curran et al, , 1992Horikami et al, 1996; Kato et al, 1997a, b;Lin et al, 2005;Malur et al, 2004;Parks et al, 2006; Patterson et al, 2000;Reutter et al, 2001;Smallwood & Moyer, 2004;Tober et al, 1998;Witko et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

The C, V and W proteins of Nipah virus inhibit minigenome replication

Sleeman

Bankamp

Hummel

et al. 2008

Journal of General Virology

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Nipah virus (NiV) is a recently emergent, highly pathogenic, zoonotic paramyxovirus of the genus Henipavirus. Like the phosphoprotein (P) gene of other paramyxoviruses, the P gene of NiV is predicted to encode three additional proteins, C, V and W. When the C, V and W proteins of NiV were tested for their ability to inhibit expression of the chloramphenicol acetyltransferase (CAT) reporter gene in plasmid-based, minigenome replication assays, each protein inhibited CAT expression in a dose-dependent manner. The C, V and W proteins of NiV also inhibited expression of CAT from a measles virus (MV) minigenome, but not from a human parainfluenzavirus 3 (hPIV3) minigenome. Interestingly, the C and V proteins of MV, which have previously been shown to inhibit MV minigenome replication, also inhibited NiV minigenome replication; however, they were not able to inhibit hPIV3 minigenome replication. In contrast, the C protein of hPIV3 inhibited minigenome replication of hPIV3, NiV and MV. Although there is very limited amino acid sequence similarity between the C, V and W proteins within the paramyxoviruses, the heterotypic inhibition of replication suggests that these proteins may share functional properties. INTRODUCTIONNipah virus (NiV) is a recently emergent, highly pathogenic, zoonotic paramyxovirus. NiV was first identified in Malaysia in 1998 during an outbreak of neurological and respiratory disease in swine, which resulted in severe febrile encephalitis in humans who had contact with infected pigs (Chua et al., 2000). NiV has been associated with subsequent outbreaks of fatal, febrile encephalitis in Bangladesh and India between (Hsu et al., 2004Luby et al., 2006). Molecular characterization of NiV revealed that it was related closely to Hendra virus, another zoonotic paramyxovirus, which had emerged in Australia in 1994 (Chua et al., 2000;Harcourt et al., 2000Harcourt et al., , 2001. These two viruses constitute the recently recognized genus Henipavirus (Mayo, 2002).The genome of NiV is 18 246 nt in length and contains six transcription units encoding six viral structural proteins (39-N-P-M-F-G-L-59) and three predicted non-structural proteins, C, V and W . As in other paramyxoviruses, the C protein of NiV is expressed from an alternative open reading frame (ORF) within the phosphoprotein (P) gene, whereas the V and W proteins are expressed by RNA editing (Lamb & Kolakofsky, 2001). At a unique, highly conserved RNA-editing site in the P gene, the polymerase protein (L) inserts a single, nontemplated G residue that results in a frame shift and the expression of the V protein. Insertion of two nontemplated G residues results in expression of the W protein . Whilst the C proteins of NiV and of other paramyxoviruses are unique and share no sequence similarity with the P protein (Lamb & Kolakofsky, 2001), the V and W proteins share an amino-terminal 407 aa domain with P and each possesses a unique carboxyl-terminal domain consisting of 52 aa for V and 47 aa for W . The sequences of the P proteins of paramyxoviruses a...

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“…The distinct cellular distributions of each protein shown in this study imply that the inhibitory effects of these proteins utilize different mechanisms. In cells infected with MeV, a specific interaction between C and L proteins is required in order to inhibit viral RNA transcription and replication, while the inhibition mediated by the V protein correlates specifically with its ability to bind RNA (Bankamp et al, 2005;Grogan & Moyer, 2001;Parks et al, 2006;Smallwood & Moyer, 2004;Witko et al, 2006). Since the W protein localizes to the nucleus, the mechanism behind its inhibition of replication is likely to be different from that of C and V, as it is unlikely that it interacts with the cytoplasmic viral polymerase.…”

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confidence: 99%

Determination of the henipavirus phosphoprotein gene mRNA editing frequencies and detection of the C, V and W proteins of Nipah virus in virus-infected cells

Harcourt²,

Mungall

et al. 2009

Journal of General Virology

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The henipaviruses, Nipah virus (NiV) and Hendra virus (HeV), are highly pathogenic zoonotic paramyxoviruses. Like many other paramyxoviruses, henipaviruses employ a process of cotranscriptional mRNA editing during transcription of the phosphoprotein (P) gene to generate additional mRNAs encoding the V and W proteins. The C protein is translated from the P mRNA, but in an alternate reading frame. Sequence analysis of multiple, cloned mRNAs showed that the mRNA editing frequencies of the P genes of the henipaviruses are higher than those reported for other paramyxoviruses. Antisera to synthetic peptides from the P, V, W and C proteins of NiV were generated to study their expression in infected cells. All proteins were detected in both infected cells and purified virions. In infected cells, the W protein was detected in the nucleus while P, V and C were found in the cytoplasm.Nipah virus (NiV) and Hendra virus (HeV) are paramyxoviruses in the genus Henipavirus, the subfamily Paramyxovirinae, within the family Paramyxoviridae. HeV causes febrile respiratory illness in humans and animals; of all the febrile illnesses caused by HeV in Australia from 1994 to 2007, there were 17 equine deaths and two human fatalities out of three cases (Hanna et al., 2006;Hooper & Williamson, 2000; Murray et al., 1995a, b). The first human NiV infections were detected during an outbreak of severe febrile encephalitis in peninsular Malaysia and Singapore from autumn 1998 to spring 1999 (Chua et al., 2000). NiV has subsequently been established as the cause of fatal human encephalitis in Bangladesh in 2001, 2003, 2007(Banerjee, 2007Hsu et al., 2004) as well as in India in (Chadha et al., 2006.Fruit-eating bats of the genus Pteropus are a natural reservoir for NiV and HeV (Chua et al., 2002;Halpin et al., 2000;Yob et al., 2001). Humans are infected by exposure to infected fruit bats or material contaminated by infected bats (Hsu et al., 2004), but are also infected via intermediate hosts such as pigs (Amal et al., 2000;Chew et al., 2000;Paton et al., 1999) or by direct human-tohuman contact (Gurley et al., 2007). No vaccines or specific antiviral drugs are currently available for NiV. Although patients from the Malaysian outbreak who received ribavirin showed a lower mortality rate (Chong et al., 2001), ribavirin was unable to protect NiV-infected hamsters from fatal disease (Georges-Courbot et al., 2006).The genomes of NiV and HeV are 18 246 and 18 234 nt, respectively, making them significantly larger than most other paramyxoviruses, with the exceptions of rodentborne Beilong and J viruses (Jack et al., 2005;Li et al., 2006). The N, P and L proteins are required and sufficient for mini-genome replication, similar to other members of the subfamily Paramyxovirinae (Halpin et al., 2004). The P genes of the subfamily Paramyxovirinae, which are 2 704 nt for NiV and 2 698 nt for HeV ( Supplementary Fig. S1, available in JGV Online), contain several open reading frames (ORFs) in addition to the P ORF. Like most other paramyxovirinae, henipavir...

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Inhibition of measles virus minireplicon-encoded reporter gene expression by V protein

Cited by 33 publications

References 58 publications

Human Parainfluenza Virus Type 2 V Protein Inhibits Genome Replication by Binding to the L Protein: Possible Role in Promoting Viral Fitness

Human Parainfluenza Virus Type 2 V Protein Inhibits Genome Replication by Binding to the L Protein: Possible Role in Promoting Viral Fitness

The C, V and W proteins of Nipah virus inhibit minigenome replication

Determination of the henipavirus phosphoprotein gene mRNA editing frequencies and detection of the C, V and W proteins of Nipah virus in virus-infected cells

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