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Papers In Press, published online ahead of print January 25, 2002
Cell Biology & Anatomy, Louisiana State University Health Sciences Center, New Orleans, LA 70112
Corresponding Author: pcserj{at}lsuhsc.edu
HAND2 is a basic helix-loop-helix (bHLH) transcription factor expressed in numerous tissues during development including the heart, limbs and a subset of neural crest derivatives. Functional analysis has shown that HAND2 is involved in development of the branchial arches, heart, limb, vasculature and nervous system. Although it is essential for development of numerous tissues, little is known about its mode of action. To address this, we have characterized HAND2 transcriptional regulatory mechanisms. Using mammalian one-hybrid analysis we show that HAND2 contains a strong transcriptional activation domain in the amino terminal third of the protein. Like most tissue-restricted bHLH factors, HAND2 heterodimerizes with the broadly expressed bHLH factors the E-proteins. We determined the consensus DNA binding site of HAND2 and show that HAND2 binds a subset of E-boxes as a heterodimer with E12. Yeast two-hybrid screening of a neuroblastoma cDNA library for HAND2 interacting proteins selected HAND2 and numerous additional members of the E-protein family. Although HAND2 homodimer formation was confirmed by in vitro analysis, HAND2 fails to homodimerize in a mammalian two-hybrid assay but demonstrates robust HAND2:E12 interaction. We conclude that HAND2 functions as a transcription activator by binding a subset of E-boxes as a heterodimer with E-proteins.
J. Biol. Chem, 10.1074/jbc.M200283200
Submitted on January 10, 2002
Revised on January 25, 2002
Accepted on January 24, 2002
The basic helix-loop-helix factor HAND2 functions as a transcriptional activator by binding to E-boxes as a heterodimer
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