JGG 2002, 29(11): 0-958 DOI:   ISSN: 1673-8527 CN: 11-5450/R           

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LIM Protein KyoT2 Interacts with Human Tight Junction Protein ZO-2-i3

HUANG Hong-Yan1;LI Rong2;SUN Qiang2; WANG Jian2; ZHOU Peng2; HAN Hua2; ZHANG Wan-Hui1

1.Department of Physiology, 2.Department of Medical Genetics and Developmental Biology, The Fourth Military Medical University, Xi’an 710032,China

Abstract

It was reported than LIM protein KyoT2 negatively regulated transcription by association with the RBP-J DNA-binding protein. Using yeast two-hybrid system with LIM protein KyoT2 as a bait, we have isolated an alternatively spliced form of human tight junction protein 2-ZO-2-i3. Sequence analysis indicated that ZO-2-i3 is composed of 19 exons, and selected usage of exons led to an alteration in the region following the kinase domain as compared with the published sequence. To identify the interaction between LIM protein KyoT2 and ZO-2-i3, yeast two-hybrid system, purification of KyoT2 protein, and GST pull-down assay were performed in the experiments. After KyoT2 and ZO-2-i3 changed vectors, positive two-hybrid yeast was obtained. Using KyoT2 protein and antibody in GST pull-down asasy positive result was also obrained. Therefore we conformed KyoT2 interacted ZO-2-i3 in vitro.Furthermore it was identified in yeast that KyoT2 associated with ZO-2-i3 through its LIM2 domain.

Keywords KyoT2   ZO-2   yeast two-hybrid system   GST pull-down  
Received 1900-01-01 Revised 1900-01-01 Online 2002-11-10 
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