STRUCTURE-FUNCTION ANALYSIS OF INTEGRASE INTERACTOR-1 HSNF5L1 REVEALSDIFFERENTIAL PROPERTIES OF 2 REPEAT MOTIFS PRESENT IN THE HIGHLY CONSERVED REGION

Retroviral integrase (IN) catalyzes the integration of retroviral cDNA into host chromosome, Ini1 (integrase interactor 1) is a host protein that specifically binds and stimulates in vitro joining activity of H IV-1 IN. Ini1 has homology to yeast transcription factor SNF5 and is a component of the analogous mammalian SWI/SNF complex that can remodel chromatin, Little is known about the function of Ini1 in mammalian ce lls, To gain insight into the functional domains of Ini1, and to under stand the details of protein-protein interactions of IN and Ini1, a st ructure-function analysis of Ini1 was initiated, By means of the yeast two-hybrid system, the minimal IN binding domain of Ini1 was characte rized, One of the two repeat motifs present in the highly conserved re gions of Ini1 was found necessary and sufficient to bind to IN in yeas t as well as in vitro, Because LN binds to only one of the two repeat motifs in this conserved region of Ini1, it appears that the IN-Ini1 i nteraction is very specific and functionally significant, Characteriza tion of DNA-binding properties of Ini1 revealed that Ini1 can bind to plasmid DNA, binding more readily to supercoiled DNA than to the relax ed circular DNA. The minimal domain for DNA binding was localized to a region upstream of repeat 1, The DNA binding activity of Ini1 is not required for its ability to interact with LN, The finding that the two repeat motifs of Ini1 display differential binding to HIV-1 IN and th at this discrete component of mammalian SWI/SNF complex binds to DNA w ill help understand the role of Ini1 in HIV-1 integration and in cellu lar process.