The Predominant Protein-arginine Methyltransferase from Saccharomyces cerevisiae

Gary, Jonathan D.; Lin, Wey Jinq; Yang, Melody C.; Herschman, Harvey R.; Clarke, Steven

doi:10.1074/jbc.271.21.12585

Cited by 190 publications

(261 citation statements)

References 67 publications

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“…Sedimentation profile of ZZ-Rrp8p, Gar1p, and large subunit ribosomal protein rpL3 on a glycerol gradient+ A total extract from strain JG590 (expressing ZZ-Rrp8p) was loaded on a 10-30% glycerol gradient and subjected to centrifugation+ Fractions were collected [numbered from 1 (top of the gradient) to 20 (bottom of the gradient)] and proteins in each fraction were precipitated with TCA, separated by SDS-PAGE, and revealed by western blotting+ Fractions containing the peak of 40S, 60S, and 80S ribosomal subunits are indicated by arrows+ which guide RNAs have been identified+ Although we deem it unlikely, the possibility remains that Rrp8p introduces some of the four 29O-ribose methylations that are unaccounted for, in particular the methylation of adenine 436 in 18S rRNA (Lowe & Eddy, 1999)+ Aside from 29-O ribose methylations, rRNAs undergo base methylations which are thought to be added by specific protein enzymes+ The only well-characterized eukaryotic rRNA base modification enzyme is Dim1p, responsible for the dimethylation of two consecutive adenines in 18S and also required for 18S rRNA synthesis (Lafontaine et al+, 1995; Lafontaine & Tollervey, 1998)+ In addition, it is likely that the nucleolar protein Nop2p, necessary for proper 27S pre-rRNA processing (Hong et al+, 1997), is a m 5 C-methyltransferase modifying rRNA (Motorin & Grosjean, 1999)+ No evidence so far allows us to exclude the possibility that Rrp8p could catalyze one or several rRNA base methylations+ Another possibility is that Rrp8p catalyzes the arginine methylation of nucleolar proteins+ Indeed it has been demonstrated that the arginine found in the FGGRGGF sequence present several times in the GAR domain is modified to asymmetric N G , N G dimethylarginine (asymmetric DMA)+ DMA can be detected FIGURE 10. Analysis of snRNAs and proteins associated with ZZ-Rrp8p+ A: Patterns of 39-end-labeled snRNAs associated with ZZ-Rrp8p+ Yeast extracts were produced from cells expressing ZZ-Nop1p (lanes 1 and 2), Gar1p-ZZ (lanes 3 and 4), or ZZ-Rrp8p (lanes 5 and 6)+ RNAs purified from the total extracts (T, lanes 1, 3, 5) or from the pellets obtained after immunoprecipitations performed in 200 mM KAc using IgG-sepharose (P, lanes 2, 4, 6) were 39 end-labeled with [ 32 P]pCp and separated on a 6% sequencing gel+ Positions of some of the C/D and H/ACA snoRNAs inferred from their size are indicated+ M: molecular weight markers (pBR322 digested with HaeIII/TaqI)+ B: Northern analysis of immunoprecipitated RNAs+ RNAs purified from pellets obtained following immunoprecipitation of Gar1p-ZZ (lane 1), ZZ-Nop1p (lane 2), or ZZ-Rrp8p from cellular extracts were separated by PAGE, transferred onto a nylon membrane, and hybridized with antisense oligodeoxynucleotide probes detecting snR10, snR42, and snR190+ C: Western analysis of proteins associated with ZZ-Rrp8p+ To attempt to characterize proteins associated with ZZ-Rrp8p, a total cellular extract from strain JG590 expressing ZZ-Rrp8p was produced using a 200-mM KAc buffer+ This extract was subjected to IgG-sepharose chromatography+ Proteins from 1/100 of total lysate (Start), 1/100 of unbound fraction (Unbound), and 1/20 of eluted fraction (Eluate) were precipitated using TCA, separated by SDS-PAGE and analyzed by western blotting+ Antibodies against U3-associated proteins (Mopp10p, Lcp5p, and Nop1p), H/ACA snoRNP proteins (Gar1p and Nhp2p), C/D snoRNP protein (Nop1p), or the nucleolar GAR domain-containing protein Nsr1p were used+ on the GAR domains of the S. cerevisiae hnRNP proteins A1, Npl3p, and Hrp1p and of the nucleolar proteins Nop1p (fibrillarin), Gar1p, and Nsr1p (Gary et al+, 1996)+ ...…”

Section: Discussionmentioning

confidence: 99%

Rrp8p is a yeast nucleolar protein functionally linked to Gar1p and involved in pre-rRNA cleavage at site A2

Bousquet‐Antonelli

Vanrobays

Gélugne

et al. 2000

RNA

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Chemical modifications and processing of the 18S, 5.8S, and 25S ribosomal RNAs from the 35S pre-ribosomal RNA depend on an important set of small nucleolar ribonucleoprotein particles (snoRNPs). Genetic depletion of yeast Gar1p, an essential common component of H/ACA snoRNPs, leads to inhibition of uridine isomerizations to pseudouridines on the 35S pre-rRNA and of the early pre-rRNA cleavages at sites A1 and A2, resulting in a loss of mature 18S rRNA synthesis. To identify Gar1p functional partners, we screened for mutations that are synthetically lethal with a gar1 mutant allele encoding a Gar1p mutant protein lacking its two glycine/arginine-rich (GAR) domains. We identified a previously uncharacterized Saccharomyces cerevisiae open reading frame, YDR083W (now designated RRP8), that encodes a highly conserved protein containing motifs found in methyltransferases. Rrp8p localizes to the nucleolus. A yeast strain lacking this protein is viable at 30 8C but displays strong growth impairment at lower temperatures. In this strain, cleavage of the pre-rRNA at site A2 is strongly affected whereas cleavages at sites A0 and A1 are only slightly inhibited or delayed.

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Section: Discussionmentioning

confidence: 99%

Rrp8p is a yeast nucleolar protein functionally linked to Gar1p and involved in pre-rRNA cleavage at site A2

Bousquet‐Antonelli

Vanrobays

Gélugne

et al. 2000

RNA

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“…3). Currently, only four PRMTs have been identified in yeast and it appears that Rmt1 (Hmt1) is the predominant arginine methyltransferase, because cells that lack Rmt1 show a significant reduction in asymmetric monomehtylation and dimethylation (Gary et al, 1996). Yeast cells also have the only type III PRMT, Rmt2, whereas Hsl7 and Skb1 represent type II PRMT in S. cerevisiae and S. pombe, respectively (Zobel-Thropp et al, 1998;Ma, 2000).…”

Section: Protein Arginine Methyltransferasesmentioning

confidence: 99%

Interplay between chromatin remodelers and protein arginine methyltransferases

Pal

Sif

2007

Journal Cellular Physiology

138

124

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Chromatin modifying enzymes have emerged as key regulators of all DNA based processes, which control cell growth, development, and differentiation. Recently, it has become clear that different chromatin remodeling and histone-modifying activities are involved in transcriptional activation and repression. Among the enzymes involved in regulating chromatin structure is the family of protein arginine methyltransferases (PRMTs) that specializes in methylating both histones as well as key cellular proteins. There are eleven different PRMT genes (PRMT1-11) whose biological function remains under explored. PRMTs regulate various cellular processes such as DNA repair and transcription, RNA processing, signal transduction, and nucleo-cytoplasmic localization. Like histone lysine methylation, methylation of histone arginine residues can either induce or inhibit transcription depending on the residue being modified and the type of methylation being introduced. In this review, we will focus on the latest findings and biological roles of ATP-dependent chromatin remodeling complexes and PRMT enzymes, and how their aberrant expression is linked to cancer.

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“…In S. cerevisiae, there are two established orthologs of the PRMT1-9 family -Rmt1 (mammalian PRMT1 ortholog; (85) and Hsl7 (mammalian PRMT5 ortholog; Ref. 86).…”

Section: An Unusual Protein Arginine Methyltransferase That Modifies mentioning

confidence: 99%

The ribosome: A hot spot for the identification of new types of protein methyltransferases

Clarke¹

2018

Journal of Biological Chemistry

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Cellular physiology depends on the alteration of protein structures by covalent modification reactions. Using a combination of bioinformatic, genetic, biochemical, and mass spectrometric approaches, it has been possible to probe ribosomal proteins from the yeast Saccharomyces cerevisiae for posttranslationally methylated amino acid residues and for the enzymes that catalyze these modifications. These efforts have resulted in the identification and characterization of the first protein histidine methyltransferase, the first N-terminal protein methyltransferase, two unusual types of protein arginine methyltransferases, and a new type of cysteine methylation. Two of these enzymes may modify their substrates during ribosomal assembly because the final methylated histidine and arginine residues are buried deep within the ribosome with contacts only with RNA. Two of these modifications occur broadly in eukaryotes, including humans, while the others demonstrate a more limited phylogenetic range. Analysis of strains where the methyltransferase genes are deleted has given insight into the physiological roles of these modifications. These reactions described here add diversity to the modifications that generate the typical methylated lysine and arginine residues previously described in histones and other proteins. Regulation of biological function by protein methylation reactionsIt is more and more apparent just how much of biological function is dependent upon posttranslational modifications (1). The human genome encodes some 900 enzymes catalyzing just protein phosphorylation or ubiquitination (2,3). However, it is now clear that methyl groups can stand beside phosphate groups and ubiquitin as major players in controlling the physiological functions of proteins. We are beginning to understand how the much greater diversity of protein methylation reactions can give rise to a greater diversity of function (1,4-8). We are also learning the importance of crosstalk between protein and DNA methylation reactions (9) and among protein methylation, phosphorylation, acetylation, and ubiquitination reactions (10-12). Finally, we are discovering how alterations in protein methylation pathways can lead to human pathology, particularly in cancer (13-15).The collection of over sixty human protein arginine and lysine methyltransferases that leave histone "marks" recognized by reader proteins to guide gene expression are perhaps the poster children for the importance of protein methyltransferases (16-17). However, recent work has emphasized the importance of methylating non-histone substrates at these residues, particularly ribosomal proteins, translation factors, and transcription factors in a wide variety of organisms (7,8,15,18,19). Furthermore, the methylation of lysine and arginine residues represents just the tip of the iceberg in protein methylation -modifications have also been established at histidine, modified histidine (diphthamide), glutamate, glutamine, asparagine, Lisoaspartate, D-aspartate, cysteine, isoprenylcysteine, me...

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The Predominant Protein-arginine Methyltransferase from Saccharomyces cerevisiae

Cited by 190 publications

References 67 publications

Rrp8p is a yeast nucleolar protein functionally linked to Gar1p and involved in pre-rRNA cleavage at site A2

Rrp8p is a yeast nucleolar protein functionally linked to Gar1p and involved in pre-rRNA cleavage at site A2

Interplay between chromatin remodelers and protein arginine methyltransferases

The ribosome: A hot spot for the identification of new types of protein methyltransferases

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