PRMT6 diminishes HIV-1 Rev binding to and export of viral RNA

Invernizzi, Cédric; Xie, Baode; Richard, Stéphane; Wainberg, Mark A.

doi:10.1186/1742-4690-3-93

Cited by 59 publications

(42 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…ity. PRMT6 has been shown to inhibit HIV-1 transcription through the methylation of Tat, Rev, and the nucleocapsid proteins (69,70,71). PRMT1 has been shown to methylate and inhibit hepatitis C virus (HCV) NS3 protein, but in turn, HCV counteracts PRMT1 repressive activity by activating protein phosphatase 2, which inhibits PRMT1 (72,73).…”

Section: Discussionmentioning

confidence: 99%

Methyltransferase PRMT1 Is a Binding Partner of HBx and a Negative Regulator of Hepatitis B Virus Transcription

Benhenda

Ducroux

Rivière

et al. 2013

J Virol

101

110

View full text Add to dashboard Cite

The hepatitis B virus X protein (HBx) is essential for virus replication and has been implicated in the development of liver cancer. HBx is recruited to viral and cellular promoters and activates transcription by interacting with transcription factors and coactivators. Here, we purified HBx-associated factors in nuclear extracts from HepG2 hepatoma cells and identified protein arginine methyltransferase 1 (PRMT1) as a novel HBx-interacting protein. We showed that PRMT1 overexpression reduced the transcription of hepatitis B virus (HBV), and this inhibition was dependent on the methyltransferase function of PRMT1. Conversely, depletion of PRMT1 correlated with increased HBV transcription. Using a quantitative chromatin immunoprecipitation assay, we found that PRMT1 is recruited to HBV DNA, suggesting a direct effect of PRMT1 on the regulation of HBV transcription. Finally, we showed that HBx expression inhibited PRMT1-mediated protein methylation. Downregulation of PRMT1 activity was further observed in HBV-replicating cells in an in vivo animal model. Altogether, our results support the notion that the binding of HBx to PRMT1 might benefit viral replication by relieving the inhibitory activity of PRMT1 on HBV transcription.

show abstract

Section: Discussionmentioning

confidence: 99%

Methyltransferase PRMT1 Is a Binding Partner of HBx and a Negative Regulator of Hepatitis B Virus Transcription

Benhenda

Ducroux

Rivière

et al. 2013

J Virol

101

110

View full text Add to dashboard Cite

show abstract

“…An important aspect of our synthesis is the use of the one-pot approach to include the desulfurization step; this reduces the number of purification steps and significantly increases the synthesis yield. Having these synthesis tools in hand, we plan to study the effect of post-translational modifications [54][55][56] on the function and structure of the Rev protein, along with introducing chemical switches [57,58] to control Rev folding and self-assembly, to better understand these processes on the function of Rev protein. [4,53,59] …”

Section: Expression Of Hiv-1 Revmentioning

confidence: 99%

Chemical Synthesis and Expression of the HIV‐1 Rev Protein

et al. 2011

View full text Add to dashboard Cite

The HIV‐1 Rev protein is responsible for shuttling partially spliced and unspliced viral mRNA out of the nucleus. This is a crucial step in the HIV‐1 lifecycle, thus making Rev an attractive target for the design of anti‐HIV drugs. Despite its importance, there is a lack of structural, biophysical, and quantitative information about Rev. This is mainly because of its tendency to undergo self‐assembly and aggregation; this makes it very difficult to express and handle. To address this knowledge gap, we have developed two new highly efficient and reproducible methods to prepare Rev in large quantities for biochemical and structural studies: 1) Chemical synthesis by using native chemical ligation coupled with desulfurization. Notably, we have optimized our synthesis to allow for a one‐pot approach for the ligation and desulfurization steps; this reduced the number of purification steps and enabled the obtaining of desired protein in excellent yield. Several challenges emerged during the design of this Rev synthesis, such as racemization, reduced solubility, formylation during thioester synthesis, and the necessity for using orthogonal protection during desulfurization; solutions to these problems were found. 2) A new method for expression and purification by using a vector that contained an HLT tag, followed by purification with a Ni column, a cation exchange column, and gel filtration. Both methods yielded highly pure and folded Rev. The CD spectra of the synthetic and recombinant Rev proteins were identical, and consistent with a predominantly helical structure. These advances should facilitate future studies that aim at a better understanding of the structure and function of the protein.

show abstract

“…PRMT6 localizes exclusively to the cell nucleus, exhibits automethylation, and methylates in vitro glycine-and arginine-rich (GAR) sequences in proteins (6). Despite its overlapping substrate specificity with PRMT1, some PRMT6-specific cellular targets do not contain the GAR consensus sequence (1,24), including high mobility group proteins HMGA1a and HMGA1b (25,26), DNA polymerase ␤ (27), HIV-1 trans-activator of transcription (Tat) protein (28), HIV-1 regulator of virion (Rev) protein (29), HIV-1 nucleocapsid protein (30), and histone H3 (31)(32)(33). Methylation of polymerase ␤ by PRMT6 was shown to increase its repair activity of damaged DNA, implicating PRMT6 as a regulator of base excision repair (27).…”

mentioning

confidence: 99%

“…Most recently several groups have demonstrated that PRMT6 methylation of Arg-2 on histone H3 in vivo exists in an exclusive relationship with Lys-4 methylation, thus adding to the mounting evidence that PRMT6 is a negative regulator of cellular as well as viral transcriptional activation (31)(32)(33). PRMT6 and other PRMTs are now a focus of attention for the development of inhibitors targeting methyl transfer activity (20,29,30,(35)(36)(37).…”

mentioning

confidence: 99%

A Kinetic Study of Human Protein Arginine N-Methyltransferase 6 Reveals a Distributive Mechanism

Lakowski

Frankel

2008

Journal of Biological Chemistry

128

View full text Add to dashboard Cite

Human protein arginine N-methyltransferase 6 (PRMT6) transfers methyl groups from the co-substrate S-adenosyl-Lmethionine to arginine residues within proteins, forming S-adenosyl-L-homocysteine as well as -N G -monomethylarginine (MMA) and asymmetric dimethylarginine (aDMA) residues in the process. We have characterized the kinetic mechanism of recombinant His-tagged PRMT6 using a mass spectrometry method for monitoring the methylation of a series of peptides bearing a single arginine, MMA, or aDMA residue. We find that PRMT6 follows an ordered sequential mechanism in which S-adenosyl-L-methionine binds to the enzyme first and the methylated product is the first to dissociate. Furthermore, we find that the enzyme displays a preference for the monomethylated peptide substrate, exhibiting both lower K m and higher V max values than what are observed for the unmethylated peptide. This difference in substrate K m and V max , as well as the lack of detectable aDMA-containing product from the unmethylated substrate, suggest a distributive rather than processive mechanism for multiple methylations of a single arginine residue. In addition, we speculate that the increased catalytic efficiency of PRMT6 for methylated substrates combined with lower K m values for native protein methyl acceptors may obscure this distributive mechanism to produce an apparently processive mechanism.Human protein arginine N-methyltransferases (PRMTs) 2 are a family of enzymes that transfer methyl groups from the co-substrate S-adenosyl-L-methionine (AdoMet) to the terminal nitrogen atoms on the guanidino groups of arginine residues within proteins, forming S-adenosyl-L-homocysteine

show abstract

PRMT6 diminishes HIV-1 Rev binding to and export of viral RNA

Cited by 59 publications

References 63 publications

Methyltransferase PRMT1 Is a Binding Partner of HBx and a Negative Regulator of Hepatitis B Virus Transcription

Methyltransferase PRMT1 Is a Binding Partner of HBx and a Negative Regulator of Hepatitis B Virus Transcription

Chemical Synthesis and Expression of the HIV‐1 Rev Protein

A Kinetic Study of Human Protein Arginine N-Methyltransferase 6 Reveals a Distributive Mechanism

Contact Info

Product

Resources

About