, 2011) and is being explored for neuroprotective therapies (Toledo et al., 2008). Thus, further study of yellow module hub proteins may yield new therapeutic targets for mitigating postsynaptic dysfunction in HD. The green module is highly enriched with proteins involved in actin cytoskeleton organization (Figure 6E). The annotation of this

module is consistent with the emerging role of Htt as a direct actin binding protein (Angeli et al., 2010), with an evolutionarily ancient function in regulating the actin-binding protein, myosin, in chemotaxis and cytokinesia (Wang et al., 2011). Green module hub proteins, including Cdk5 (Anne et al., 2007), Rph3A (Smith et al., 2007), Rock2 (Shao et al., 2008), and Gja1/Connexin-43 (Vis et al., 1998), have previously been implicated in HD. Therefore, this module reveals a set of actin binding candidates that can be studied in the context of normal Htt function and HD pathogenesis. www.selleckchem.com/products/isrib-trans-isomer.html Finally, the cyan module is the only age-dependent module and is also highly enriched with proteins residing in mitochondria, or functioning in inflammation, G protein signaling and as modifiers of mHtt aggregation. Several of these proteins (Usp9x, Rock1, and Sirt2) appear to modify mHtt aggregation or toxicity in cellular models and are known selleck chemical drug targets (Kaltenbach et al., 2007, Shao et al.,

2008, Pallos et al., 2008 and Luthi-Carter et al., 2010). The role of cyan module proteins in influencing mHtt aggregation gained further support after a genome-wide RNA interference screen for modifiers of mHtt aggregation identified a number of similar genes, including Aldoa, Csnk2a1, Hspa9, Pfkm, and Rab1a in Drosophila cells ( Zhang et al.,

2010). Thus, the cyan module is enriched with proteins complexed with Htt in an age-dependent manner, which may help to probe the poorly understood role of aging in the pathogenesis of HD. We next sought to validate those proteins not previously implicated in HD but with a high red module connectivity (hub proteins) as Htt physical interactors and/or genetic modifiers (Table 1). We performed an anti-Htt Linifanib (ABT-869) co-IP from BACHD and WT mouse brains followed by western blot analysis for the specific candidate proteins to confirm interaction (Figure 7A). We confirmed six proteins (Atp1a1, Atp2b2, Cct8, Cct1, Tuba1b, and Cntn1) and one positive control red module protein (Hap40/F8A1) as coimmunoprecipitating with Htt. Furthermore, Vps35, a protein in the retromer complex that functions in the retrieval of certain membrane proteins from the endosome to the plasma membrane (Attar and Cullen, 2010), was found in both the IP of Htt in both the BACHD and WT mouse brain extracts (Figure 7B). Moreover, Htt is also present in the reciprocal co-IP of Vps35 from these brain extracts (Figure 7B). Thus, the top red module proteins are indeed complexed with Htt in the mouse brain.