Characterization of p73 functional domains necessary for transactivation and growth suppression.
p73, a p53 family member, is highly similar to p53 in both structure and function. Like p53, the p73 protein contains an N-terminal activation domain, a DNA-binding domain, a tetramerization domain, and several PXXP motifs. Previously, we and others have shown that some functional domains in p53, such as the DNA-binding and tetramerization domains, are required for inducing both cell cycle arrest and apoptosis whereas others, such as the second activation domain, the proline-rich domain, and the C-terminal basic domain, are only required for inducing apoptosis. To determine the activity of p73 functional domains, we have generated stable inducible cell lines that express p73beta and various mutants deficient in one or more functional domains. We found that in addition to the DNA-binding domain, p73-mediated growth suppression requires the N-terminal activation domain and the tetramerization domain. However, unlike p53, p73-mediated apoptosis does not require the region adjacent to the activation domain or the entire C-terminal region. Interestingly, while the N- or the C-terminal PXXP motifs are dispensable for p73 function, deletion of both the N- and the C-terminal PXXP motifs renders p73 inactive in transactivation. In addition, we found that substitution of two conserved tandem hydrophobic residues with two hydrophilic ones, which can abrogate the activity of the first activation domain in p53, has no effect on p73 transcriptional activity. Together, we showed that the p73 protein has its own unique determinants for transactivation and growth suppression.
Nozell
Susan
S
Department of Cell Biology, The University of Alabama at Birmingham, Birmingham, AL 35294-0005, USA.
Wu
Yijun
Y
McNaughton
Kelly
K
Liu
Gang
G
Willis
Amy
A
Paik
Jason C
JC
Chen
Xinbin
X
D020816:Amino Acid Motifs; D017209:Apoptosis; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology; D016147:Genes, Tumor Suppressor; D006131:Growth Inhibitors / Q000737:chemistry* / Q000502:physiology; D006801:Humans; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins / Q000737:chemistry* / Q000502:physiology; D015533:Transcriptional Activation; D000072160:Tumor Protein p73; D025521:Tumor Suppressor Proteins
D004268:DNA-Binding Proteins; D006131:Growth Inhibitors; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; C000606460:TP73 protein, human; D000072160:Tumor Protein p73; D025521:Tumor Suppressor Proteins