Nucleic Acids Research, 1988, Vol. 16, No. 8 3239-3253
© 1988
Articles |
In vivo functional analysis of in vitro protein binding sites in the immunoglobulin heavy chain enhancer
1Department of Biological Chemistry, UCLA School of Medicine, University of California Los Angeles, CA 90024, USA 2Molecular Biology Institute, University of California Los Angeles, CA 90024, USA
Received January 14, 1988. Revised March 18, 1988. Accepted March 18, 1988.
We have systematically investigated the functional role of protein binding sites within the mouse immunoglobulin heavy chain enhancer which we previously identified by in vitro binding studies (1,2). Each binding site was deleted, mutant enhancers were cloned 3' of the chloramphenicol acetyl transferase gene in the vector pA10CAT2 and transfected into plasmacytoma cells. We demonstrate that the newly identified site E, located at 324–338 bp, is important for enhancer function; previously identified sites B(uE1), C1(uE2), C2(uE3) and C3 were also shown to be important for enhancer activity. Sites A and D are not required for IgH enhancer function, as assayed by our methods. Thus, including the octamer site, six protein binding sites which bind at least six different proteins are important for enhancer function in vivo.
*Present address: Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142, USA
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