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PMID:16923123
| Citation |
Itoh, T, Satoh, M, Kanno, E and Fukuda, M (2006) Screening for target Rabs of TBC (Tre-2/Bub2/Cdc16) domain-containing proteins based on their Rab-binding activity. Genes Cells 11:1023-37 |
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| Abstract |
It has recently been proposed that the TBC (Tre2/Bub2/Cdc16) domain functions as a GAP (GTPase-activating protein) domain for small GTPase Rab. Because of the large number of Rab proteins in mammals, however, most TBC domains have never been investigated for Rab-GAP activity. In this study we established panels of the GTP-fixed form of 60 different Rabs constructed in pGAD-C1, a yeast two-hybrid bait vector. We also constructed a yeast two-hybrid prey vector (pGBDU-C1) that harbors the cDNA of 40 distinct TBC proteins. Systematic investigation of 2400 combinations of 60 GTP-fixed Rabs and 40 TBC proteins by yeast two-hybrid screening revealed that seven TBC proteins specifically and differentially interact with specific Rabs (e.g. OATL1 interacts with Rab2A; FLJ12085 with Rab5A/B/C; and Evi5-like with Rab10). Measurement of in vitro Rab-GAP activity revealed that OATL1 and Evi5-like actually possess significant Rab2A- and Rab10-GAP activity, respectively, but that FLJ12085 do not display Rab5A-GAP activity at all. These results indicate that specific interaction between TBC protein and Rab would be a useful indicator for screening for the target Rabs of some TBC/Rab-GAP domains, but that there is little correlation between the Rab-binding activity and Rab-GAP activity of other TBC proteins. |
| Links |
PubMed Online version:10.1111/j.1365-2443.2006.00997.x |
| Keywords |
Animals; COS Cells; Cell Cycle Proteins/chemistry; Cell Cycle Proteins/metabolism; Cercopithecus aethiops; GTPase-Activating Proteins/metabolism; Humans; Mice; Molecular Sequence Data; NIH 3T3 Cells; Nuclear Proteins/metabolism; Phylogeny; Protein Binding; Protein Structure, Tertiary; Transcription Factors/metabolism; Two-Hybrid System Techniques; rab GTP-Binding Proteins/metabolism |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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