Nuclear export of hnRNP Hrp1p and nuclear export of hnRNP Npl3p are linked and influenced by the methylation state of Npl3p.
Eukaryotic mRNA processing and export are mediated by a series of complexes composed of heterogeneous nuclear ribonucleoproteins (hnRNPs). Many of these hnRNPs are methylated at arginine residues within their RGG domains. Although cellular arginine methylation is required for the efficient nuclear export of several hnRNPs, its role in this process is unknown. To address this question, we replaced the methylated RGG tripeptides of two hnRNPs, Npl3p and Hrp1p, with KGG. We found that these substitutions specifically abolish their methylation but have different effects on their nuclear export activity. Although the efficient export of Hrp1p requires cellular methyltransferase activity, the modification of Hrp1p itself is dispensable. In contrast, we found that Npl3 arginine methylation not only facilitates its own export but also is required for Hrp1p to efficiently exit the nucleus. Consistent with this observation, we found that Npl3p and Hrp1p exist in a ribonucleoprotein complex. We provide the first evidence that the arginine methylation of a particular protein directly affects its activity. Efficient export does not require methylation per se, but unmethylated arginine residues lead to retention of hnRNPs. Thus, arginine methylation serves to mask the Npl3p RGG domain for efficient ribonucleoprotein export.
Xu
Chong
C
Department of Molecular Biology, UMNDJ-SOM, 2 Medical Center Drive, Stratford, NJ 08084, USA.
Henry
Michael F
MF
D021581:Active Transport, Cell Nucleus; D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism*; D000483:Alleles; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001120:Arginine / Q000378:metabolism; D001345:Autoradiography; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism*; D004265:DNA Helicases / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry; D005456:Fluorescent Dyes; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D017930:Genes, Reporter; D034441:Heterogeneous-Nuclear Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007211:Indoles; D008745:Methylation; D008780:Methyltransferases / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009154:Mutation; D011233:Precipitin Tests; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D014316:Tritium
D005456:Fluorescent Dyes; D005656:Fungal Proteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D007211:Indoles; D014316:Tritium; C007293:DAPI; D001120:Arginine; D008780:Methyltransferases; D000251:Adenosine Triphosphatases; D004265:DNA Helicases