Hazzard for figure assistance.

J.A. was supported by National Eye Institute (NEI)/National Institutes of Health (NIH) grants R01EY018350, R01EY018836, R01EY020672, R01EY022238, R21EY019778, RC1EY020442, Doris Duke selleckchem Distinguished Clinical Scientist Award, Burroughs Wellcome Fund Clinical Scientist Award in Translational Research, Dr. E. Vernon Smith and Eloise C. Smith Macular Degeneration Endowed Chair, and B.J.F. was supported by by NIH T32HL091812 and UL1RR033173. J.A. is named as an inventor on patent applications about age-related macular degeneration filed by the University of Kentucky and is a founder of iVeena Pharmaceuticals, which is commercializing these technologies. “
“Huntington’s disease (HD) is one of the most common dominantly inherited neurodegenerative disorders clinically characterized by a triad of movement disorder, cognitive dysfunction, and psychiatric impairment (Bates et al., 2002). HD neuropathology is characterized by selective and massive degeneration of the striatal medium spiny neurons (MSNs) and, to a lesser extent, the deep layer cortical pyramidal neurons (Vonsattel and DiFiglia, 1998). The disease is caused by a CAG repeat expansion resulting in an elongated polyglutamine (polyQ) stretch near the N terminus of Huntingtin (Htt) (The Huntington’s

Disease Collaborative Research Group, 1993). HD is one of nine polyQ disorders with shared molecular genetic features, such as an inverse relationship between the expanded repeat length and the age of disease onset, and evidence for toxic gain-of-function as a result of Selleck Rucaparib the polyQ expansion (Orr and Zoghbi, 2007). However, each of the polyQ disorders appears to target a distinct subset of neurons

in the brain Megestrol Acetate leading to disease-specific symptoms. Hence, it is postulated that molecular determinants beyond the polyQ repeat itself may be critical to disease pathogenesis (Orr and Zoghbi, 2007). Protein-interacting cis-domains ( Lim et al., 2008) and posttranslational modifications (PTMs) of polyQ proteins ( Emamian et al., 2003 and Gu et al., 2009) can significantly modify disease pathogenesis in vivo. Thus, studying the proteins that interact with domains beyond the polyQ region may provide important clues to disease mechanisms. In the case of HD, several hundred putative Htt interactors have been discovered using ex vivo methods, such as yeast two-hybrid (Y2H) or in vitro affinity pull-down assays, utilizing only small N-terminal fragments of Htt ( Goehler et al., 2004 and Kaltenbach et al., 2007). Such studies have provided insight into Htt’s normal function as a scaffolding protein involved in vesicular and axonal transport and nuclear transcription ( Caviston and Holzbaur, 2009 and Li and Li, 2006). The caveats of the prior Htt interactome studies include the exclusive use of small Htt N-terminal fragments as baits and the isolation of interactors ex vivo.