SLX5 and Huntington’s Disease: Second Update

In my last blog post, I spoke about developing a protocol to verify findings from a new data set. This has been my focus since that update.

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June Update: Change of Plans

Upon returning to campus to begin my project, I found out that new data related to my project had been found while I had been away. This new data identified 398 genes whose transcription was affected (either up or down regulated) by aggregated Huntingtin’s protein (Htt). Also included was the reversal of the negative effects of Htt when a gene known as Slx5 was over-expressed. There is a lot of background information required to understand why these findings are so important.

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Abstract: Identifying Genes that Counteract Huntington’s Disease Pathology

Researching the ways harmful aggregated proteins, which cause many neurodegenerative diseases, are rendered toxic in basic research is an important first step in finding cures for these diseases. This study looks specifically at the nuclear activities of part of the gene which leads to the production of the mutant protein (Htt) which cause Huntington’s disease. Inside the cell mutant htt forms protein aggregates, and also enters the nucleus where it misregulates the activity (transcription) of genes. This inappropriate transcriptional regulation is believed to cause severe stress to cells. Previous research in the Kerscher lab found that certain proteins could reduce Huntington’s inappropriate transcriptional activity, and decrease its negative effects on the cell. The protein they identified is involved in protein degradation; this discovery lead to the question of whether other proteins are also capable of decreasing the transcriptional promiscuity of the Huntington’s protein. In this project, many proteins will be tested using a novel assay to identify proteins that are capable of decreasing the negative effects of the Huntington’s protein. This approach is called a genetic screen. To do this, a technique will be developed prior to the study, in which a yeast strain with a reporter gene will be transformed into yeast cells expressing mutant huntingtin to reveal the typical transcriptional promiscuity of the huntingtin protein. The production of this reporter will be assayed on growth media that may contains chromogenic substrates. For example, one reporter gene I could use is Mel 1. Colonies expressing a Mel 1 reporter construct will be blue in color, allowing us to easily recognize them. In this study, suppressor genes will then be transformed into the cells, to identify those that show a decrease in reporter gene activity, depending on the assay used. A decrease in reporter activty would indicate suppression of huntingtin’s transcriptional activity by a particular suppressor. I will isolate and sequence this suppressor and functionally analyze it.  Since most yeast genes are conserved in humans this work may lead us to important novel insights about similar mechanisms in human cells that can counteract the progression of Huntington’s Disease.