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(American Journal of Pathology. 2003;163:453-464.)
© 2003 American Society for Investigative Pathology

Transgenic Mice Demonstrate Novel Promoter Regions for Tissue-Specific Expression of the Urokinase Receptor Gene

Heng Wang*, John Hicks{dagger}, Parham Khanbolooki*, Sun-Jin Kim*, Chunhong Yan*, Yao Wang{ddagger} and Douglas Boyd*

From the Department of Cancer Biology,*MD Anderson Cancer Center, Houston, Texas; the Department of Pathology,{dagger}Texas Children’s Hospital, Houston, Texas; and Orthopaedic Research Institute,{ddagger}St. George Hospital Clinical School, the University of New South Wales, Kogarah, Sydney, Australia

The urokinase-type plasminogen activator receptor (u-PAR) contributes to cell migration and proteolysis in normal and cancerous tissues. Currently, there are no reports on the regulatory regions directing tissue-specific expression. Consequently, we undertook a study to identify novel promoter regions required for expression of this gene in transgenic mice bearing a LacZ reporter regulated by varying amounts (0.4, 1.5, and 8.5 kb) of upstream sequence. The 0.4-kb u-PAR upstream sequence directed weak and strong LacZ expression in the placenta and epididymis, respectively, both of which are tissues that express endogenous u-PAR. Conversely, transgene expression in the apical cells of the colon positive for endogenous u-PAR protein required 1.5 kb of upstream sequence for optimal expression. Furthermore, chromatin accessibility assays coupled with real-time polymerase chain reaction suggested a putative regulatory region spanning -1295/-1192 driving u-PAR expression in colonic cells. Interestingly, placental transgene expression was augmented with the 8.5-kb upstream fragment compared with the shorter 1.5-kb fragment indicating contributing element(s) between -1.5 and -8.5 kb. Thus, while 0.4 kb of upstream sequence directs u-PAR expression in the epididymis, sequences located between -0.4 and -1.5 kb and between -1.5 and -8.5 kb are required for optimal tissue-specific expression in the colon and the placenta, respectively.




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H. Wang, L. Yang, Md. S. Jamaluddin, and D. D. Boyd
The Kruppel-like KLF4 Transcription Factor, a Novel Regulator of Urokinase Receptor Expression, Drives Synthesis of This Binding Site in Colonic Crypt Luminal Surface Epithelial Cells
J. Biol. Chem., May 21, 2004; 279(21): 22674 - 22683.
[Abstract] [Full Text] [PDF]


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