PRMT8, a New Membrane-bound Tissue-specific Member of the Protein Arginine Methyltransferase Family
Protein arginine methylation is a common post-translational modification that has been implicated in signal transduction, RNA processing, transcriptional regulation, and DNA repair. A search of the human genome for additional members of the protein arginine N-methyltransferase (PRMT) family of enzymes has identified a gene on chromosome 12 that we have termed PRMT8. This novel enzyme is most closely related to PRMT1, although it has a distinctive N-terminal region. The unique N-terminal end harbors a myristoylation motif, and we have shown here that PRMT8 is indeed modified by the attachment of a myristate to the glycine residue after the initiator methionine. The myristoylation of PRMT8 results in its association with the plasma membrane. The second singular property of PRMT8 is its tissue-specific expression pattern; it is largely expressed in the brain. A glutathione S-transferase fusion protein of PRMT8 has type I PRMT activity, catalyzing the formation of ω-NG-monomethylated and asymmetrically ω-NG,NG-dimethylated arginine residues on a recombinant glycine- and arginine-rich substrate. PRMT8 is thus an active arginine methyltransferase that is membrane-associated and tissue-specific, two firsts for this family of enzymes. Protein arginine methylation is a common post-translational modification that has been implicated in signal transduction, RNA processing, transcriptional regulation, and DNA repair. A search of the human genome for additional members of the protein arginine N-methyltransferase (PRMT) family of enzymes has identified a gene on chromosome 12 that we have termed PRMT8. This novel enzyme is most closely related to PRMT1, although it has a distinctive N-terminal region. The unique N-terminal end harbors a myristoylation motif, and we have shown here that PRMT8 is indeed modified by the attachment of a myristate to the glycine residue after the initiator methionine. The myristoylation of PRMT8 results in its association with the plasma membrane. The second singular property of PRMT8 is its tissue-specific expression pattern; it is largely expressed in the brain. A glutathione S-transferase fusion protein of PRMT8 has type I PRMT activity, catalyzing the formation of ω-NG-monomethylated and asymmetrically ω-NG,NG-dimethylated arginine residues on a recombinant glycine- and arginine-rich substrate. PRMT8 is thus an active arginine methyltransferase that is membrane-associated and tissue-specific, two firsts for this family of enzymes. Arginine methylation is a widespread post-translational modification that is mediated by the protein arginine methyltransferase (PRMT) 4The abbreviations used are: PRMT, protein arginine N-methyltransferase; ADMA, asymmetric ω-NG,NG-dimethylarginine; CARM1, coactivator-associated arginine methyltransferase 1; GST, glutathione S-transferase; GAR, glycine/arginine-rich; GST-GAR, GST fusion protein containing the GAR N-terminal region of fibrillarin; GFP, green fluorescent protein; FLAG, peptide epitope DYKDDDDK; PVDF, polyvinylidene fluoride; EST, expressed sequence tag.4The abbreviations used are: PRMT, protein arginine N-methyltransferase; ADMA, asymmetric ω-NG,NG-dimethylarginine; CARM1, coactivator-associated arginine methyltransferase 1; GST, glutathione S-transferase; GAR, glycine/arginine-rich; GST-GAR, GST fusion protein containing the GAR N-terminal region of fibrillarin; GFP, green fluorescent protein; FLAG, peptide epitope DYKDDDDK; PVDF, polyvinylidene fluoride; EST, expressed sequence tag. family of enzymes (1Gary J.D. Clarke S. Prog. Nucleic Acid Res. Mol. Biol. 1998; 61: 65-131Crossref PubMed the family of protein arginine two type I enzymes that and asymmetrically arginine residues and type enzymes that and arginine PRMT1, and type I and is a type The post-translational methylation of arginine residues has been shown to protein by and Clarke S. S. Biol. PubMed S. S. Mol. PubMed Biol. PubMed Arginine methylation is in a of DNA RNA signal transduction, protein and protein S. Mol. PubMed PRMT to identified and the enzyme methyltransferase J.D. Clarke S. Biol. PubMed Mol. Biol. PubMed identified the type I enzyme J.D. Clarke S. Biol. PubMed of its to arginine 1998; PubMed for this enzyme has to a in a J.D. Clarke S. Biol. 1998; PubMed identified in a an enzyme that the PubMed a protein S. Biol. PubMed identified by to members of this enzyme family Clarke S. Biol. PubMed in a for that to a S. Res. of PRMT1, and a J.D. Clarke S. Biol. 1998; PubMed Clarke S. Biol. PubMed S. Clarke S. Biol. PubMed S. Clarke S. Biol. PubMed of a GAR motif, and the methylation PubMed PubMed is that the type I and type enzymes of the This has been shown for the S. Clarke S. Res. PubMed the S. Mol. PubMed and for S. S. Mol. Biol. PubMed PRMT that and residues the on the arginine residue asymmetrically we the of the PRMT PRMT8. This enzyme type I and is to a GAR The sequence of PRMT8 is to expressed PRMT1, PRMT8 is expressed in the brain. unique to PRMT8 is an N-terminal that harbors a myristoylation and a of the of this PRMT8 a of we used the sequence to for used to PRMT8 by the and The with and and used to and by and by with the and The with and and a modified The modified a and and a The and the and The with and and and by human with DNA to the PRMT8 the fusion by enzyme the human DNA used a DNA with a Clarke S. Biol. PubMed and have been Clarke S. Biol. 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PubMed PubMed A search of the human and expressed sequence a novel PRMT that has been This PRMT has been in the for and The sequence has a This enzyme a of with the arginine methyltransferase The of this has been PubMed PubMed although the unique N-terminal region of This N-terminal region is and and an in PubMed This enzyme has been termed PRMT8 S. Mol. PubMed S. and we this of DNA the human and genome that and PRMT8 on is on chromosome and is on chromosome and in to the region of chromosome 12 and is on chromosome The of PRMT8 and in and a of an in the expression of RNA a of human PRMT8 expression in the a of and two of and This has been with Clarke S. Biol. PubMed this we a that to the of after with PRMT8 The of the PRMT8 with is the RNA a of identified for and human and of that the of the a in that used and a in a PRMT8 identified in and the of identified PRMT8 a that is expressed in S. S. 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PubMed PubMed The and PubMed PubMed in the PRMT8 and the recombinant of GAR and thus is a type I protein arginine recombinant PRMT8 and arginine with a of in the of for in a of of The GST, GST-GAR, and and with and for with for and on a with and methylation in on in the of the substrate. asymmetric and with to and of to of and of for the type of of for methyltransferase a substrate. the formation of and thus that PRMT8 is a type I is for PRMT8 The that the of and PRMT8 the that PRMT8 in to PRMT1, it I a have been to the and the of the and in the and the Clarke S. Biol. PubMed S. PubMed and in of Clarke S. Biol. PubMed S. Biol. 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PubMed PRMT8 harbors a myristoylation that is for its we a A in The is to the end of thus the for The and a by a the N-terminal end of the fusion protein its to with the plasma and a to of PRMT8 its plasma The and and and and by with with GFP, and and with for The fusion with by to a with and to for after the to with to the of of fusion protein in that PRMT8 is indeed the with GFP, and and with for The fusion by to a and to is and the of the glycine residue to an residue results in a of PRMT8 is a and with arginine PRMT1, methyltransferase and PubMed Biol. PubMed S. S. PubMed a the region. of methyltransferase is is PubMed Biol. 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PubMed PRMT8 an in the of a and gene to the of PRMT8 in this of PRMT8 is to PRMT1, with enzymes the of and and to PRMT1, PRMT8 type I PRMT8 with and to the plasma membrane. the of two a of by the is to PRMT8 that and A to methylation is a of protein S. Mol. PubMed of the and human that most have been an gene on human that is most closely related to This gene is a for and its expression it is that we have the in this we to and of arginine Arginine methylation is a widespread post-translational modification that is mediated by the protein arginine methyltransferase (PRMT) 4The abbreviations used are: PRMT, protein arginine N-methyltransferase; ADMA, asymmetric ω-NG,NG-dimethylarginine; CARM1, coactivator-associated arginine methyltransferase 1; GST, glutathione S-transferase; GAR, glycine/arginine-rich; GST-GAR, GST fusion protein containing the GAR N-terminal region of fibrillarin; GFP, green fluorescent protein; FLAG, peptide epitope DYKDDDDK; PVDF, polyvinylidene fluoride; EST, expressed sequence tag.4The abbreviations used are: PRMT, protein arginine N-methyltransferase; ADMA, asymmetric ω-NG,NG-dimethylarginine; CARM1, coactivator-associated arginine methyltransferase 1; GST, glutathione S-transferase; GAR, glycine/arginine-rich; GST-GAR, GST fusion protein containing the GAR N-terminal region of fibrillarin; GFP, green fluorescent protein; FLAG, peptide epitope DYKDDDDK; PVDF, polyvinylidene fluoride; EST, expressed sequence tag. family of enzymes (1Gary J.D. Clarke S. Prog. Nucleic Acid Res. Mol. Biol. 1998; 61: 65-131Crossref PubMed the family of protein arginine two type I enzymes that and asymmetrically arginine residues and type enzymes that and arginine PRMT1, and type I and is a type The post-translational methylation of arginine residues has been shown to protein by and Clarke S. S. Biol. PubMed S. S. Mol. PubMed Biol. PubMed Arginine methylation is in a of DNA RNA signal transduction, protein and protein S. Mol. PubMed The PRMT to identified and the enzyme methyltransferase J.D. Clarke S. Biol. PubMed Mol. Biol. PubMed identified the type I enzyme J.D. Clarke S. Biol. PubMed of its to arginine 1998; PubMed for this enzyme has to a in a J.D. Clarke S. Biol. 1998; PubMed identified in a an enzyme that the PubMed a protein S. Biol. PubMed identified by to members of this enzyme family Clarke S. Biol. PubMed in a for that to a S. Res. The of PRMT1, and a J.D. Clarke S. Biol. 1998; PubMed Clarke S. Biol. PubMed S. Clarke S. Biol. PubMed S. Clarke S. Biol. PubMed of a GAR motif, and the methylation PubMed PubMed is that the type I and type enzymes of the This has been shown for the S. Clarke S. Res. PubMed the S. Mol. PubMed and for S. S. Mol. Biol. PubMed PRMT that and residues the on the arginine residue asymmetrically we the of the PRMT PRMT8. This enzyme type I and is to a GAR The sequence of PRMT8 is to expressed PRMT1, PRMT8 is expressed in the brain. unique to PRMT8 is an N-terminal that harbors a myristoylation and a of the of this PRMT8 a of we used the sequence to for used to PRMT8 by the and The with and and used to and by and by with the and The with and and a modified The modified a and and a The and the and The with and and and by human with DNA to the PRMT8 the fusion by enzyme the human DNA used a DNA with a Clarke S. Biol. PubMed and have been Clarke S. Biol. 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PubMed PubMed A search of the human and expressed sequence a novel PRMT that has been This PRMT has been in the for and The sequence has a This enzyme a of with the arginine methyltransferase The of this has been PubMed PubMed although the unique N-terminal region of This N-terminal region is and and an in PubMed This enzyme has been termed PRMT8 S. Mol. PubMed S. and we this of DNA the human and genome that and PRMT8 on is on chromosome and is on chromosome and in to the region of chromosome 12 and is on chromosome The of PRMT8 and in and a of an in the expression of RNA a of human PRMT8 expression in the a of and two of and This has been with Clarke S. Biol. PubMed this we a that to the of after with PRMT8 The of the PRMT8 with is the RNA a of identified for and human and of that the of the a in that used and a in a PRMT8 identified in and the of identified PRMT8 a that is expressed in S. S. PubMed This of to by used a of RNA for and PRMT8 identified of that expressed in of that PRMT8 is largely an enzyme of the PRMT8 expression in and in the of PRMT8 and PRMT8 expression and identified it an I Protein Arginine arginine have been shown to type I activity, results in the of and enzymes J.D. Clarke S. Biol. PubMed J.D. Clarke S. Biol. 1998; PubMed PubMed and Clarke S. Biol. PubMed has been shown for S. Clarke S. Biol. PubMed and this for S. Biol. PubMed has been shown to a to arginine residues Clarke S. Biol. PubMed PRMT8 is active type of it we a fusion and its methyltransferase a of The GST-GAR, and by PRMT1, and J.D. Clarke S. Biol. PubMed Clarke S. Biol. PubMed PubMed The and PubMed PubMed in the PRMT8 and the recombinant of GAR and thus is a type I protein arginine recombinant PRMT8 and arginine with a of in the of for in a of of The GST, GST-GAR, and and with and for with for and on a with and methylation in on in the of the substrate. asymmetric and with to and of to of and of for the type of of for methyltransferase a substrate. the formation of and thus that PRMT8 is a type I is for PRMT8 The that the of and PRMT8 the that PRMT8 in to PRMT1, it I a have been to the and the of the and in the and the Clarke S. Biol. PubMed S. PubMed and in of Clarke S. Biol. PubMed S. Biol. 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Biol. PubMed Clarke S. Mol. PubMed for this of enzymes Clarke S. Biol. PubMed PubMed A search of the human and expressed sequence a novel PRMT that has been This PRMT has been in the for and The sequence has a This enzyme a of with the arginine methyltransferase The of this has been PubMed PubMed although the unique N-terminal region of This N-terminal region is and and an in PubMed This enzyme has been termed PRMT8 S. Mol. PubMed S. and we this of DNA the human and genome that and PRMT8 on is on chromosome and is on chromosome and in to the region of chromosome 12 and is on chromosome The of PRMT8 and in and a of an PRMT8 in the expression of RNA a of human PRMT8 expression in the a of and two of and This has been with Clarke S. Biol. PubMed this we a that to the of after with PRMT8 The of the PRMT8 with is the RNA a of identified for and human and of that the of the a in that used and a in a PRMT8 identified in and the of identified PRMT8 a that is expressed in S. S. PubMed This of to by used a of RNA for and PRMT8 identified of that expressed in of that PRMT8 is largely an enzyme of the PRMT8 expression in and in the of PRMT8 and PRMT8 expression and identified it an PRMT8 I Protein Arginine arginine have been shown to type I activity, results in the of and enzymes J.D. Clarke S. Biol. PubMed J.D. Clarke S. Biol. 1998; PubMed PubMed and Clarke S. Biol. PubMed has been shown for S. Clarke S. Biol. PubMed and this for S. Biol. PubMed has been shown to a to arginine residues Clarke S. Biol. PubMed PRMT8 is active type of it we a fusion and its methyltransferase a of The GST-GAR, and by PRMT1, and J.D. Clarke S. Biol. PubMed Clarke S. Biol. PubMed PubMed The and PubMed PubMed in the PRMT8 and the recombinant of GAR and thus the type of of for methyltransferase a substrate. the formation of and thus that PRMT8 is a type I is for PRMT8 The that the of and PRMT8 the that PRMT8 in to PRMT1, it I PRMT8 a have been to the and the of the and in the and the Clarke S. Biol. PubMed S. PubMed and in of Clarke S. Biol. PubMed S. Biol. PubMed the of we it to the N-terminal end of to its of to PRMT1, we to a of to with and with A the plasma membrane-associated of a to the of the and the N-terminal PRMT8 to the on the plasma by a of a region that has an and is in residues PubMed the of the protein to a that is the PubMed the of a protein that the PubMed of the PRMT8 sequence a myristoylation the unique N-terminal end of the This S. PubMed is and and the PubMed is a of to this that with the and to the association of PRMT8. with the of a myristate to the N-terminal glycine residue after the of the methionine. a glycine to the myristoylation PubMed S. PubMed PRMT8 harbors a myristoylation that is for its we a A in The is to the end of thus the for The and a by a the N-terminal end of the fusion protein its to with the plasma and a to that PRMT8 is indeed the with GFP, and and with for The fusion by to a and to is and the of the glycine residue to an residue results in a of PRMT8 is a PRMT8 and with arginine PRMT1, methyltransferase and PubMed Biol. PubMed S. S. PubMed a the region. of methyltransferase is is PubMed Biol. 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PubMed PRMT8 an in the of a and gene to the of PRMT8 in this of PRMT8 is to PRMT1, with enzymes the of and and to PRMT1, PRMT8 type I PRMT8 with and to the plasma membrane. the of two a of by the is to PRMT8 that and A to methylation is a of protein S. Mol. PubMed of the and human that most have been an gene on human that is most closely related to This gene is a for and its expression it is that we have the in this we to and of arginine A of that and PRMT8 the of this enzyme PRMT8 has in and PubMed a PRMT8 is in the is an This that PRMT8 a gene of an gene after the of the PRMT8 has a unique N-terminal region that on it the to with the plasma The of a has that PRMT8 has an property that the PRMT to to the plasma PRMT8 is the of this family of enzymes that a tissue-specific expression The largely expression of PRMT8 that it a in protein methylation shown to a in Biol. PubMed Clarke S. Res. PubMed it that methyltransferase with signal This with the that methyltransferase Biol. PubMed and that is after the of with S. Res. PubMed PRMT8 an in the of a and gene to the of PRMT8 in this The of PRMT8 is to PRMT1, with enzymes the of and and to PRMT1, PRMT8 type I PRMT8 with and to the plasma membrane. the of two a of by the is to PRMT8 that and A to Arginine methylation is a of protein S. Mol. PubMed of the and human that most have been an gene on human that is most closely related to This gene is a for and its expression it is that we have the in this we to and of arginine The of for with the and members of the and Clarke for this