Expression in a RabGAP yeast mutant of two human homologues, one of which is an oncogene.
The yeast proteins Msb3p and Msb4p are two Ypt/Rab-specific GTPase-activating proteins (GAPs) involved in cell growth polarization. Both proteins share with a wide variety of other proteins the highly conserved TBC domain forming the catalytically active RabGAP domain. In particular, Msb3p and Msb4p are similar to the human proteins oncTre210p (the 786-amino-acid product of the human Tre2 oncogene, implicated in Ewing's sarcoma) and RN-tre (a Rab5-GAP controlling endocytosis of the EGFR). To further understand the biochemical function of Tre2 oncogene, we expressed its cDNA and, as a control, the RN-tre cDNA, in an msb3 msb4 double mutant yeast strain. Complementation data show that RN-tre can, unlike Tre2, replace the function of the MSB3 and MSB4 genes. As two highly conserved amino acids, including the catalytic arginine, are mutated in the oncTre210p TBC domain, we restored these two amino acids and expressed the modified Tre2 cDNA in the yeast mutant.
Bizimungu
Christelle
C
Animal and Microbial Biology Unit, Gembloux Agricultural University, B-5030 Gembloux, Belgium. bizimungu.c@fsagx.ac.be
De Neve
Nancy
N
Burny
Arsène
A
Bach
Stéphane
S
Bontemps
Françoise
F
Portetelle
Daniel
D
Vandenbol
Micheline
M
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D001665:Binding Sites; D020134:Catalytic Domain; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D020690:GTPase-Activating Proteins / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D005816:Genetic Complementation Test; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D009154:Mutation; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015514:Oncogene Proteins, Fusion / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D043222:Ubiquitin Thiolesterase; D020691:rab GTP-Binding Proteins / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D020690:GTPase-Activating Proteins; D047908:Intracellular Signaling Peptides and Proteins; C076360:MSB2 protein, S cerevisiae; C540987:MSB3 protein, S cerevisiae; C540988:MSB4 protein, S cerevisiae; D015513:Oncogene Proteins; D015514:Oncogene Proteins, Fusion; D011518:Proto-Oncogene Proteins; D029701:Saccharomyces cerevisiae Proteins; C100665:USP6NL protein, human; D010450:Endopeptidases; C473330:USP6 protein, human; D043222:Ubiquitin Thiolesterase; D020691:rab GTP-Binding Proteins