SPIN6 is a Rho GTPase-activating protein (RhoGAP) and interacts with SPL11 <i>in vitro</i> and <i>in vivo</i>.

<p>(A) The protein structure of SPIN6, SPIN6.2 and SPIN6.3. The position of the Pleckstrin Homology (PH) domain (34–142), RhoGAP domain (192–383), and coiled coil (CC) motif (583–699) are indicated. (B) SPIN6 interacts with SPL11 in yeast. SPL11 represents the full-length SPL11; ARM is the ARM domain of SPL11; and SPL11m is the SPL11 mutant with a three amino-acid deletion at C314P315T316 in the U-box domain, resulting in a loss-of-function of E3 ligase activity (Zeng et al., 2004). After they were diluted 10, 100, and 1000 times with sterilized-distilled H₂O, the Mav203 yeast transformants were plated on synthetic dextrose medium without Trp, Leu, and His amino acids (SD-LTH) and with 0 mM or 40 mM 3-amino-1,2,4,-triazole (3AT), separately. (C) SPIN6 binds to SPL11 in the GST pull-down assay. His:SPIN6 and GST:SPL11, GST:ARM of SPL11 were used in the assay. (D) SPIN6 interacts with SPL11 in <i>N</i>. <i>benthamiana</i> in BiFC assay. SPIN6 was fused with the C-terminal of eYFP to make SPIN6:CeYFP. SPL11, ARM, and SPL11m were fused with the N-terminal eYFP to make NeYFP:SPL11, NeYFP:ARM, and NeYFP:SPL11m, respectively.