Methods to Discover Alternative Promoter Usage and Transcriptional Regulation of Murine Bcrp1.
Gene expression in different tissues is often controlled by alternative promoters that result in the synthesis of mRNA with unique - usually untranslated - first exons. Bcrp1 (Abcg2), the murine orthologue of the ABC transporter Breast Cancer Resistance Protein (BCRP, ABCG2), has at least four alternative promoters that are designated by the corresponding four alternative first exons produced: E1U, E1A, E1B, and E1C. Herein, in-silico protocols are presented to predict alternative promoter usage for Bcrp1. Furthermore, reporter assay methods are described to produce reporter constructs for alternative promoters and to determine the functionality of putative promoters upstream of the alternative first exons that are identified.
Greenebaum Cancer Center, University of Maryland School of Medicine; Pharmaceutical Sciences, University of Maryland School of Pharmacy.
Karthika
K
Natarajan
Greenebaum Cancer Center, University of Maryland School of Medicine; Baltimore VA Medical Center.
Yi
Y
Xie
Membrane Transport and Biopharmaceutics, School of Pharmaceutical Sciences, Kanazawa University.
Takeo
T
Nakanishi
Obstetrics, Gynecology and Reproductive Science, University of Pittsburgh; Magee Women's Research Institute.
Rebecca S
RS
Moreci
Obstetrics, Gynecology, Perinatal Research Branch (NICHD), Wayne State University School of Medicine.
Pancharatnam
P
Jeyasuria
Greenebaum Cancer Center, University of Maryland School of Medicine; Baltimore VA Medical Center; Medicine, University of Maryland School of Medicine; Pathology, University of Maryland School of Medicine.
Arif
A
Hussain
Greenebaum Cancer Center, University of Maryland School of Medicine; Baltimore VA Medical Center; Medicine, University of Maryland School of Medicine; Pathology, University of Maryland School of Medicine; Pharmacology, University of Maryland School of Medicine; Experimental Therapeutics, University of Maryland School of Medicine; DRoss@som.umaryland.edu.
Douglas D
DD
Ross
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2 / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D005091:Exons; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; C501412:Abcg2 protein, mouse; D012333:RNA, Messenger