The yeast ribosome synthesis factor Emg1 is a novel member of the superfamily of alpha/beta knot fold methyltransferases.
Emg1 was previously shown to be required for maturation of the 18S rRNA and biogenesis of the 40S ribosomal subunit. Here we report the determination of the crystal structure of Emg1 at 2 A resolution in complex with the methyl donor, S-adenosyl-methionine (SAM). This structure identifies Emg1 as a novel member of the alpha/beta knot fold methyltransferase (SPOUT) superfamily. In addition to the conserved SPOUT core, Emg1 has two unique domains that form an extended surface, which we predict to be involved in binding of RNA substrates. A point mutation within a basic patch on this surface almost completely abolished RNA binding in vitro. Three point mutations designed to disrupt the interaction of Emg1 with SAM each caused>100-fold reduction in SAM binding in vitro. Expression of only Emg1 with these mutations could support growth and apparently normal ribosome biogenesis in strains genetically depleted of Emg1. We conclude that the catalytic activity of Emg1 is not essential and that the presence of the protein is both necessary and sufficient for ribosome biogenesis.
Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, UMR8619, Bât 430, Université de Paris-Sud, 91405 Orsay Cedex, France. nicolas.leulliot@u-psud.fr
Nicolas
N
Leulliot
Markus T
MT
Bohnsack
Marc
M
Graille
David
D
Tollervey
Herman
H
Van Tilbeurgh
D000595:Amino Acid Sequence; D001665:Binding Sites; D020134:Catalytic Domain; D019281:Dimerization; D008780:Methyltransferases / Q000737:chemistry* / Q000145:classification / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D012331:RNA, Fungal / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000145:classification / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000145:classification / Q000378:metabolism; D016415:Sequence Alignment
C444457:EMG1 protein, S cerevisiae; D012331:RNA, Fungal; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; D012436:S-Adenosylmethionine; D008780:Methyltransferases