Defining the regulatory elements in the proximal promoter of DeltaNp63 in keratinocytes: Potential roles for Sp1/Sp3, NF-Y, and p63.
p63, a homolog of the tumor suppressor p53, plays an important role in the formation of stratified epithelium such as those in the epidermis of the skin. The p63 gene gives rise to multiple functionally distinct protein isoforms, including the DeltaNp63 class of isoforms, which lacks the N-terminal transactivation domain and is synthesized from an internal promoter. DeltaNp63 proteins are the predominant isoforms expressed in keratinocytes and are thought to be important for maintenance of the proliferative capacity of these cells. Here, we have examined the transcriptional control mechanisms that govern the expression DeltaNp63 in keratinocytes. We first performed DNase I hypersensitive site mapping and demonstrated that the promoter region of DeltaNp63 is in open chromatin state in keratinocytes. To identify the cis-elements that regulate DeltaNp63, we have performed transient transfection assays in keratinocytes with several DeltaNp63 promoter constructs. This identified a short evolutionarily conserved fragment that harbors most of the transcriptional activity of the DeltaNp63 promoter. Biochemical studies of this element have revealed critical roles for CCAAT-box-binding factor (CBF/NF-Y) and Sp1/Sp3 family of proteins. In addition, our data suggest that DeltaNp63 is recruited to and can activate its own promoter, possibly through protein-protein interactions, thus providing an auto-regulatory loop of self-regulation. These studies support the notion that unique and distinct pathways control the expression of individual p53 family members and their various isoforms.
Romano
Rose-Anne
RA
Department of Biochemistry, State University of New York at Buffalo, Buffalo, New York, USA.
Birkaya
Barbara
B
Sinha
Satrajit
S
D000818:Animals; D001483:Base Sequence; D023081:CCAAT-Binding Factor / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002843:Chromatin / Q000737:chemistry; D004247:DNA / Q000032:analysis / Q000235:genetics; D004252:DNA Mutational Analysis; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D015603:Keratinocytes / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D010750:Phosphoproteins / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D048328:Potoroidae; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology; D011485:Protein Binding; D020033:Protein Isoforms / Q000032:analysis / Q000502:physiology; D050436:Regulatory Elements, Transcriptional / Q000235:genetics* / Q000502:physiology; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000502:physiology*; D051705:Sp3 Transcription Factor / Q000235:genetics / Q000502:physiology*; D015534:Trans-Activators / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D014162:Transfection
D023081:CCAAT-Binding Factor; D002843:Chromatin; D010750:Phosphoproteins; D020033:Protein Isoforms; D016329:Sp1 Transcription Factor; C497122:Sp3 protein, mouse; D015534:Trans-Activators; C486419:Trp63 protein, mouse; D051705:Sp3 Transcription Factor; D004247:DNA