Home LiteratureArticle Details

PMID: 16923123 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Screening for target Rabs of TBC (Tre-2/Bub2/Cdc16) domain-containing proteins based on their Rab-binding activity.

Genes to cells : devoted to molecular & cellular mechanisms ·Vol. 11 ·No. 9 ·2006-09-00 ·Pages 1023-37

Itoh T, Satoh M, Kanno E, Fukuda M

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.

MeSH Terms

Animals COS Cells Cell Cycle Proteins/chemistry,metabolism Chlorocebus 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

Chemicals

Cell Cycle Proteins EVI5 protein, human Evi5 protein, mouse GTPase-Activating Proteins Nuclear Proteins Transcription Factors rab GTP-Binding Proteins

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Itoh Takashi

Fukuda Initiative Research Unit, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

Satoh Megumi

Kanno Eiko

Fukuda Mitsunori

Article Info

Journal

Genes to cells : devoted to molecular & cellular mechanisms

Abbr.

Genes Cells

ISSN

1356-9597

DOI

10.1111/j.1365-2443.2006.00997.x

Published

2006-09-00

Pages

1023-37

Language

English

Region

England

NLM ID

9607379

Subset

Databases

GENBANK

AB232583, AB232584, AB232585, AB232586, AB232587, AB232588, AB232589, AB232590, AB232591, AB232592, AB232593, AB232594, AB232595, AB232596, AB232597, AB232598, AB232599, AB232600, AB232601, AB232602, AB232603, AB232604, AB232605, AB232606, AB232607, AB232608, AB232609, AB232610, AB232611, AB232612, AB232613, AB232614, AB232615, AB232616, AB232617, AB232618, AB232619, AB232620, AB232621, AB232622, AB232623, AB232624, AB232625, AB232626, AB232627, AB232628, AB232629, AB232630, AB232631, AB232632, AB232633, AB232634, AB232635, AB232636, AB232637, AB232638, AB232639, AB232640, AB232641, AB232642

External Links

PubMed source DOI: 10.1111/j.1365-2443.2006.00997.x View journal details

Analysis Services

Screening for target Rabs of TBC (Tre-2/Bub2/Cdc16) domain-containing proteins based on their Rab-binding activity.

Contact

WeChat Official Account