CLAR1, A Novel Gene Associated with Prostate Tumor Progression

摘要

The purpose of our research was to characterize the function of the novel gene Clar1 as it relates to the progression of human prostate cancer. The scope of this work included generating antibodies to Clar1, their use in characterizing the level of this protein ill prostate cancer, examining the relationship of Clar1 protein expression to tumor progression, and determining the biological function of Clar1. We have successfully developed, tested, and utilized antibodies to the Clar1 protein to characterize Clar1 expression in prostate cancer tissues. We have characterized Clar1 expression in variety of human tissues using western analysis and demonstrated predominantly cytoplasmic staining of the protein by immunohistochemistry (IHC). We have also performed IHC analysis on tissue sections obtained from 23 different prostate tumors and we are currently completing IHC on an additional 27 samples. In the 23 samples analyzed we found that 65% of the prostate cancers exhibited some level of Clarl expression. We have created Clar1 plasmid constructs for use as 'bait' proteins to identify possible Clar1 binding partners. We employed a modified yeast two- hybrid, dual bait procedure that minimized non-specific protein-protein interactions. Using this method we have identified beta-tubulin 2 as a binding partner for the Clar1 SH3 domains, suggesting that Clar1 may participate as an adaptor protein at the cell surface, and participate in membrane modifications and or signal transduction.