Molecular Recognition in Biological Systems and Bioinformatics

Transcription regulation; Developmental Biology; Mouse knockouts; Genetics; Biochemistry; Transgenics

The epidermis is stratified squamous epithelium found at the skin surface. The basal cells of the epidermis tucked away in the innermost layer constitute the proliferative compartment of epidermis. In a balanced and carefully orchestrated differentiation program, basal cells periodically withdraw from the cell cycle and migrate towards the skin surface. As these cells differentiate, they progress through distinct stages and unique sets of genes are turned on and off. Temporally and spatially compartmentalized, the epidermis thus provides a unique system to study the molecular switches that control tissue-specific and differentiation-specific gene expression. Study in our laboratory focuses on the identification and characterization of transcription factors that govern this program of control of the developmental and differentiation decisions of the epidermal cells.

1. The first area of interest of the lab includes analysis of promoters/enhancers of those genes that are unique to the various differentiation stages of the epidermis. These studies involve DNAse I hypersensitive site mapping, gel-shifts, footprints, ChIP (chromatin immunoprecipitation) assays, keratinocyte cell culture and transgenic mice analysis. To complement these studies we are also using genomic tools and in silico analysis to characterize putative cis-elements common for epidermal specific genes.

2. The second area of interest centers on families of transcription factors that play a key role in governing keratinocyte-specific gene expression. We have generated transgenic mice where the expression and function of three families of transcription factors (p63, AP-2 and Ets) can be altered in the skin epidermis in an inducible fashion. Alterations in the transcriptional regulation program in these transgenic mice leads to dramatic effects on cell development and differentiation decisions that control epidermis and skin appendages such as hair follicles and sebaceous glands. We have utilized these in vivo models to identify and characterize the molecular mechanisms that dictate normal epithelial cell fate of the skin and to gain new insights into how normal control processes are broken down during cancer initiation and progression. Our future goal is to identify the target genes for these factors and to gain insights into the process of how epidermal-specific patterns of gene expression are established.

Mouse, human

• P63
• Ets
• AP-2
• Basonuclin

Skin diseases and Cancer

• gene targeting
• Transgenics
• transient and stable transfection
• QPCR
• EMSA
• ChIP
• Histology

• HeLa
• Epidermal keratinocyte
• oral keratinocyte
• PTK2
• ES cells
• MCF
• HC11
• Cos

Wound healing, skin cancer, breast cancer

Skin and mammary Gland

http://www.smbs.buffalo.edu/bch/Labs/SinhaLab/index.htm