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. 2006 Sep;74(9):5362–5373. doi: 10.1128/IAI.00539-06

FIG. 4.

FIG. 4.

Rab4 interacts with C. trachomatis CT229 in a guanine nucleotide-dependent manner. (A) S. cerevisiae strain AH109 containing pGBKT7, pGBKBD-p53, or the indicated constitutively active pGBK-Rab constructs was mated with S. cerevisiae strain Y187 expressing pGADT7, pGAD-ct229 (101-215), or pGAD-ct229 (101-197). To identify possible interacting clones, diploids, which were selected on plates lacking Leu and Trp, were diluted into sterile water and transferred to plates lacking Leu, Trp, His, and Ade using a multipronged liquid transfer tool. GAL4AD-CT229 (101-215), but not GAL4AD-CT229 (101-197), interacted with only GAL4BD-Rab4AQ67L and not any of the other Rab GTPases tested. (B) S. cerevisiae strain Y187 expressing either pGADT7, pGAD-ct229 (101-215), or pGAD-ct229 (101-197) was mated with S. cerevisiae strain AH109 expressing pGBK-Rab4A, pGBK-Rab4AQ67L, and pGBK-Rab4AS22N. Diploids were selected and screened as described for panel A. GAL4AD-CT229 (101-215) interacted with constitutively active GAL4BD-Rab4AQ67L, and to a lesser extent with wild-type GAL4BD-Rab4A (data not shown), but not with inactive GDP-restricted GAL4BD-Rab4AS22N.