This project will characterize the minimal promoter region of human NAT1 and NAT2 and study effects of genetic polymorphisms on expression in human mammary cells.
Specific Aim 1 involves identifying the NAT1 and NAT2 transcriptional start site in human mammary epithelial cells by 5'-RLM-RACE.
Specific Aims 2 and 3 will identify the functional promoter of NAT1 and NAT2 by construction of luciferase reporter genes containing 5'-UTR regions of NAT1 and AT2 and site-directed mutagenesis and deletion analysis, and assessing the transcriptional effects of these regions by transient expression in MCF-7 and MCF-10A cell lines.
Specific Aim 4 involves investigating transcriptional regulation of NAT1 and NAT2 in normal human mammary tissues by determining NAT1 and NAT2 mRNA levels, protein expression, intrinsic protein stability, and catalytic activity for N- and O-acetylation activities as a function of 5'-UTR and coding region genotypes.
Specific Aim 5 involves sequencing the 5'-UTR of NAT1 and NAT2 in human mammary tissues to identify and characterize effects of polymorphisms in the 5'-UTR of NAT1 and NAT2. Knowledge of transcriptional regulation in the 5'-UTR of NAT1 and NAT2 will allow for an increased understanding the role of genetic polymorphisms towards genetic predisposition to breast cancer from environmental carcinogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Individual Predoctoral NRSA for M.D./Ph.D. Fellowships (ADAMHA) (F30)
Project #
5F30ES012557-03
Application #
6906583
Study Section
Special Emphasis Panel (ZRG1-F09 (20))
Program Officer
Humble, Michael C
Project Start
2003-07-15
Project End
2007-07-14
Budget Start
2005-07-15
Budget End
2006-07-14
Support Year
3
Fiscal Year
2005
Total Cost
$46,485
Indirect Cost
Name
University of Louisville
Department
Pharmacology
Type
Schools of Medicine
DUNS #
057588857
City
Louisville
State
KY
Country
United States
Zip Code
40292
Husain, Anwar; Zhang, Xiaoyan; Doll, Mark A et al. (2007) Functional analysis of the human N-acetyltransferase 1 major promoter: quantitation of tissue expression and identification of critical sequence elements. Drug Metab Dispos 35:1649-56
Husain, Anwar; Loehle, Jennifer A; Hein, David W (2007) Clinical pharmacogenetics in pediatric patients. Pharmacogenomics 8:1403-11
Husain, Anwar; Zhang, Xiaoyan; Doll, Mark A et al. (2007) Identification of N-acetyltransferase 2 (NAT2) transcription start sites and quantitation of NAT2-specific mRNA in human tissues. Drug Metab Dispos 35:721-7
Barker, David F; Husain, Anwar; Neale, Jason R et al. (2006) Functional properties of an alternative, tissue-specific promoter for human arylamine N-acetyltransferase 1. Pharmacogenet Genomics 16:515-25
Husain, Anwar; Barker, David F; States, J Christopher et al. (2004) Identification of the major promoter and non-coding exons of the human arylamine N-acetyltransferase 1 gene (NAT1). Pharmacogenetics 14:397-406