Interaction of hTCF4 by yeast two-hybrid system

To investigate the interaction of hTCF4, the yeast two-hybrid system has be en used for testing the interaction of mutants of hTCF4 with themselves. Mu tants of hTCF4(hTCF4 I) and hTCF4 II) have been obtained by polymerase chai n 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 reve rse yeast two-hybrid system (GIBCOBRL Co.). However, no interactions are fo und in hTCF4 II with hTCF4 I and in hTCF4 I with hTCF4 I. These results sug gest that hTCF4 could interact with itself to form homodimer or homocopolym er and perform the transcriptional activating function through LZ or HLH mo tifs in nucleus of renal cell carcinoma.