title: Identification of HYPK-interacting proteins reveals involvement of HYPK in regulating cell growth, cell cycle, unfolded protein response and cell death.
abstract: Huntingtin Yeast Two-Hybrid Protein K (HYPK) is an intrinsically unstructured huntingtin (HTT)-interacting protein with chaperone-like activity. To obtain more information about the function(s) of the protein, we identified 27 novel interacting partners of HYPK by pull-down assay coupled with mass spectrometry and, further, 9 proteins were identified by co-localization and co-immunoprecipitation (co-IP) assays. In neuronal cells, (EEF1A1 and HSPA1A), (HTT and LMNB2) and (TP53 and RELA) were identified in complex with HYPK in different experiments. Various Gene Ontology (GO) terms for biological processes, like protein folding (GO: 0006457), response to unfolded protein (GO: 0006986), cell cycle arrest (GO: 0007050), anti-apoptosis (GO: 0006916) and regulation of transcription (GO: 0006355) were significantly enriched with the HYPK-interacting proteins. Cell growth and the ability to refold heat-denatured reporter luciferase were decreased, but cytotoxicity was increased in neuronal cells where HYPK was knocked-down using HYPK antisense DNA construct. The proportion of cells in different phases of cell cycle was also altered in cells with reduced levels of HYPK. These results show that HYPK is involved in several biological processes, possibly through interaction with its partners.
authors.affiliation: Crystallography and Molecular Biology Division, Saha Institute of Nuclear Physics, Kolkata, West Bengal, India.
authors.forename: Kamalika Roy
authors.initials: KR
authors.lastname: Choudhury
authors.forename: Swasti
authors.initials: S
authors.lastname: Raychaudhuri
authors.forename: Nitai P
authors.initials: NP
authors.lastname: Bhattacharyya
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle; D016923:Cell Death; D049109:Cell Proliferation; D002470:Cell Survival; D019295:Computational Biology / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D000071058:Huntingtin Protein; D007150:Immunohistochemistry / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D011485:Protein Binding; D017510:Protein Folding; D050486:Serotonin Plasma Membrane Transport Proteins / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D056811:Unfolded Protein Response
chemical_list: D002352:Carrier Proteins; C086055:HTT protein, human; C526912:HYPK protein, human; C581869:HYPK protein, mouse; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D050486:Serotonin Plasma Membrane Transport Proteins; C493231:Slc6a4 protein, mouse