The intrinsic GTPase activity of the Gtr1 protein from Saccharomyces cerevisiae

Sökning: L773:1471 2091 > The intrinsic GTPas...

Sengottaiyan, PalaniveluLinnéuniversitetet,Institutionen för naturvetenskap, NV,Bengt Persson,School of Natural Sciences, Linnaeus University, Kalmar, Sweden (författare)

BackgroundThe Gtr1 protein of Saccharomyces cerevisiae is a member of the RagA subfamily of the Ras-like small GTPase superfamily. Gtr1 has been implicated in various cellular processes. Particularly, the Switch regions in the GTPase domain of Gtr1 are essential for TORC1 activation and amino acid signaling [R. Gong, L. Li, Y. Liu, P. Wang, H. Yang, L. Wang, J. Cheng, K.L. Guan, Y. Xu, Genes Dev. 25 (2011) 1668–1673]. Therefore, knowledge about the biochemical activity of Gtr1 is required to understand its mode of action and regulation.ResultsBy employing tryptophan fluorescence analysis and radioactive GTPase assays, we demonstrate that Gtr1 can adopt two distinct GDP- and GTP-bound conformations, and that it hydrolyses GTP much slower than Ras proteins. Using cysteine mutagenesis of Arginine-37 and Valine-67, residues at the Switch I and II regions, respectively, we show altered GTPase activity and associated conformational changes as compared to the wild type protein and the cysteine-less mutant.ConclusionsThe extremely low intrinsic GTPase activity of Gtr1 implies requirement for interaction with activating proteins to support its physiological function. These findings as well as the altered properties obtained by mutagenesis in the Switch regions provide insights into the function of Gtr1 and its homologues in yeast and mammals.

NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
NATURVETENSKAP Biologi hsv//swe
NATURAL SCIENCES Biological Sciences hsv//eng
Gtr1
GTPase
Intrinsic tryptophan fluorescence
Rag GTPase
Cysteine mutagenesis
Switch region
Biokemi
Biochemistry
Gtr1
GTPase
Intrinsic tryptophan fluorescence
Rag GTPase
Cysteine
mutagenesis
Switch region
Cysteine mutagenesis

Spetea, Cornelia,1968Gothenburg University,Göteborgs universitet,Institutionen för biologi och miljövetenskap,Department of Biological and Environmental Sciences(Swepub:gu)xspeco (författare)
Lagerstedt, JensLund University,Lunds universitet,Cellulär biomekanik,Forskargrupper vid Lunds universitet,Cellular Biomechanics,Lund University Research Groups(Swepub:lu)med-jls (författare)
Samyn, Dieter R.Linnéuniversitetet,Institutionen för naturvetenskap, NV,Bengt Persson,School of Natural Sciences, Linnaeus University, Kalmar, Sweden(Swepub:oru)drsn (författare)
Andersson, Michael R.Linnéuniversitetet,Institutionen för naturvetenskap, NV,Bengt Persson,School of Natural Sciences, Linnaeus University, Kalmar, Sweden(Swepub:lnu)eanmi (författare)
Ruiz-Pavon, LorenaLinnéuniversitetet,Institutionen för naturvetenskap, NV,Bengt Persson,School of Natural Sciences, Linnaeus University, Kalmar, Sweden(Swepub:lnu)erulo (författare)
Persson, Bengt L.Linnéuniversitetet,Institutionen för naturvetenskap, NV,Katholieke Universiteit Leuven, Belgium ; Flanders Institute of Biotechnology, Belgium,Bengt Persson,School of Natural Sciences, Linnaeus University, Kalmar, Sweden; Department of Molecular Microbiology, Institute of Botany and Microbiology, Katholieke Universiteit Leuven, Leuven-Heverlee, Flanders, Belgium(Swepub:lnu)hpebe (författare)
LinnéuniversitetetInstitutionen för naturvetenskap, NV (creator_code:org_t)

Av författaren/redakt...: Sengottaiyan, Pa ...; Spetea, Cornelia ...; Lagerstedt, Jens; Samyn, Dieter R.; Andersson, Micha ...; Ruiz-Pavon, Lore ...; visa fler...; Persson, Bengt L ...; visa färre...

Om ämnet

Av lärosätet: Linnéuniversitetet; Örebro universitet; Lunds universitet; Göteborgs universitet

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