Interaction of hTCF4 by yeast two-hybrid system

To investigate the interaction of hTCF4, the yeast two-hybrid system has been used for testing the interaction of mutants of hTCF4 with themselves. Mutants of hTCF4 (hTCF4 1 ) and hTCF4 II) have been obtained by polymerase chain reaction (PCR). Bait (hTCF4 I -pDBLeu and hTCF4 II -pDBLeu) and prey (hTCF4 I -pPC86 and hTCF4 II -pPC86) have been constructed by DNA recombination for yeast two-hybrid. Interaction of hTCF4 II with itself is found in the reverse yeast two-hybrid system (GIBCOBRL Co.). However, no interactions are found in hTCF4 II with hTCF4 1 and in hTCF4 1 with hTCF4 1. These results suggest that hTCF4 could interact with itself to form homodimer or homocopolymer and perform the transcriptional activating function through LZ or HLH motifs in nucleus of renal cell carcinoma.