Isolation of mouse THP gene promoter and demonstration of its kidney-specific activity in transgenic mice.
Tamm-Horsfall protein (THP), the most abundant urinary protein synthesized by the kidney epithelial cells, is believed to play important and diverse roles in the urinary system, including renal water balance, immunosuppression, urinary stone formation, and inhibition of bacterial adhesion. In the present study, we describe the isolation of a 9.3-kb, 5'-region of the mouse THP gene and show the highly conserved nature of its proximal 589-bp, 5'-flanking sequence with that in rats, cattle, and humans. We also demonstrate using the transgenic mouse approach that a 3.0-kb, proximal 5'-flanking sequence is sufficient to drive the kidney-specific expression of a heterologous reporter gene. Within the kidney, transgene expression was confined to the renal tubules that endogenously expressed the THP protein, which suggests specific transgene activity in the thick ascending limb of the loop of Henle and early distal convoluted tubules. Our results establish the kidney- and nephron-segment-specific expression of the mouse THP gene. The availability of the mouse THP gene promoter that functions in vivo should facilitate additional studies of the molecular mechanisms of kidney-specific gene regulation and should provide new molecular tools for better understanding renal physiology and disease through nephron-specific gene targeting.
Zhu
Xinhua
X
Department of Urology, Kaplan Comprehensive Cancer Center, New York University School of Medicine, New York 10016, USA.
Cheng
Jin
J
Gao
Jing
J
Lepor
Herbert
H
Zhang
Zhong-Ting
ZT
Pak
Joanne
J
Wu
Xue-Ru
XR
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins; D007668:Kidney / Q000502:physiology*; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009088:Mucoproteins / Q000235:genetics*; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes / Q000235:genetics; D058949:Uromodulin
D008164:Luminescent Proteins; D009088:Mucoproteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C548612:UMOD protein, human; C548613:Umod protein, mouse; D058949:Uromodulin; D049452:Green Fluorescent Proteins