dc.creator | Graindorge, J. S. | en |
dc.creator | Senger, B. | en |
dc.creator | Tritch, D. | en |
dc.creator | Simos, G. | en |
dc.creator | Fasiolo, F. | en |
dc.date.accessioned | 2015-11-23T10:29:08Z | |
dc.date.available | 2015-11-23T10:29:08Z | |
dc.date.issued | 2005 | |
dc.identifier | 10.1021/bi049024z | |
dc.identifier.issn | 62960 | |
dc.identifier.uri | http://hdl.handle.net/11615/28181 | |
dc.description.abstract | Yeast methionyl-tRNA synthetase (MetRS) and glutamyl-tRNA synthetase (GluRS) possess N-terminal extensions that bind the cofactor Arc1p in trans. The strength of GluRS-Arc1p interaction is high enough to allow copurification of the two macromolecules in a 1:1 ratio, in contrast to MetRS. Deletion analysis from the C-terminal end of the GluRS appendix combined with previous N-terminal deletions of GluRS allows restriction of the Arc1p binding site to the 110-170 amino acid region of GluRS. This region has been shown to correspond to a novel protein-protein interaction domain present in both GluRS and Arc1p but not in MetRS [Galani, K., Grosshans, H., Deinert, K., Hurt, E. C., and Simos, G. (2001) EMBO J. 20, 6889-6898]. The GluRS apoenzyme fails to show significant kinetics of tRNA aminoacylation and charges unfractionated yeast tRNA at a level 10-fold reduced compared to Arc1p-bound GluRS. The Km values for tRNA Glu measured in the ATP-PPi exchange were similar for the two forms of GluRS, whereas kcat is increased 2-fold in the presence of Arc1p. Band-shift analysis revealed a 100-fold increase in tRNA binding affinity when Arc1p is bound to GluRS. This increase requires the RNA binding properties of the full-length Arc1p since Arc1p N domain leaves the K d of GluRS for tRNA unchanged. Transcripts of yeast tRNA Glu were poor substrates for measuring tRNA aminoacylation and could not be used to clarify whether Arc1p has a specific effect on the tRNA charging reaction. | en |
dc.source.uri | http://www.scopus.com/inward/record.url?eid=2-s2.0-13444302274&partnerID=40&md5=0ad72e29823b3b0925615ae60987a1ab | |
dc.subject | Acylation | en |
dc.subject | Adenosinetriphosphate | en |
dc.subject | Amino acids | en |
dc.subject | Macromolecules | en |
dc.subject | Modulation | en |
dc.subject | Purification | en |
dc.subject | RNA | en |
dc.subject | Substrates | en |
dc.subject | Yeast | en |
dc.subject | Aminoacylation | en |
dc.subject | Band-shift analysis | en |
dc.subject | Binding properties | en |
dc.subject | Yeast glutamyl-tRNA synthetase (GLuRS) | en |
dc.subject | Enzymes | en |
dc.subject | amino acid transfer RNA ligase | en |
dc.subject | glutamic acid transfer RNA | en |
dc.subject | analytic method | en |
dc.subject | animal cell | en |
dc.subject | article | en |
dc.subject | binding site | en |
dc.subject | enzyme activation | en |
dc.subject | enzyme activity | en |
dc.subject | gene deletion | en |
dc.subject | genetic analysis | en |
dc.subject | macromolecule | en |
dc.subject | nonhuman | en |
dc.subject | priority journal | en |
dc.subject | protein analysis | en |
dc.subject | protein folding | en |
dc.subject | protein function | en |
dc.subject | protein protein interaction | en |
dc.subject | Adenosine Triphosphate | en |
dc.subject | Amino Acid Sequence | en |
dc.subject | Base Sequence | en |
dc.subject | Diphosphates | en |
dc.subject | Gene Expression Regulation, Fungal | en |
dc.subject | Glutamate-tRNA Ligase | en |
dc.subject | Kinetics | en |
dc.subject | Molecular Sequence Data | en |
dc.subject | Peptide Fragments | en |
dc.subject | Protein Binding | en |
dc.subject | Protein Structure, Tertiary | en |
dc.subject | RNA, Fungal | en |
dc.subject | RNA, Transfer, Glu | en |
dc.subject | RNA-Binding Proteins | en |
dc.subject | Saccharomyces cerevisiae | en |
dc.subject | Saccharomyces cerevisiae Proteins | en |
dc.subject | Transcription, Genetic | en |
dc.subject | Animalia | en |
dc.title | Role of Arc1p in the modulation of yeast glutamyl-tRNA synthetase activity | en |
dc.type | journalArticle | en |