Fusion of Skp1 and NLS to TIR1 enhances growth defects of mcm4-aid in the presence of auxin. (A) Schematic illustrations of AtTIR1 genes fused with NLS and Skp1. Two copies of NLS sequences derived from SV40 were inserted between AtTIR1 and 9myc tag (upper), while fission yeast Skp1 (SpSkp1) with 16-amino-acid linker peptides (see Methods) was fused to AtTIR1 at the N-terminus (lower). (B) Serially diluted cultures of HM1910 mcm4-aid leu1-32 cells carrying the pREP41D vector alone (MKF53), pREP41D-AtTIR1 (MKF54), pREP41D-AtTIR1-NLS (MKF55) and pREP41D-Skp1-AtTIR1 (MKF60) were spotted onto EMM plates containing auxin (0.5 mM) and incubated at 25°C. (C) Schematic illustration of the Skp1-AtTIR1- NLS-9myc fusion is presented.
Kanke et al. BMC Cell Biology 2011 12:8 doi:10.1186/1471-2121-12-8