Proteomic Analysis of Wild-type and Mutant Huntingtin-associated Proteins in Mouse Brains Identifies Unique Interactions and Involvement in Protein Synthesis

Culver, Brady P.; Savas, Jeffrey N.; Park, Sung K.; Choi, Jeong H.; Zheng, Shaoyong; Zeitlin, Scott; Yates, John R.; Tanese, Naoko

doi:10.1074/jbc.m112.359307

“…Using subcellular fractionation of brain homogenates, one study found molecular chaperones as mutant HTT interactors, in support of previous findings and highlighting the central role of these folding machineries as HTT modifiers [112]. Interestingly, components of the translational machinery were also identified as mutant HTT interacting proteins, providing a novel molecular pathway for HTT-mediated toxicity.…”

Section: Proteomic Screeningsupporting

confidence: 63%

“…Recently, proteomic studies to identify HTT-associated proteins have also been done in the context of full-length HTT expressed in mouse brain, using affinity purificationmass spectrometry [112,113]. Using subcellular fractionation of brain homogenates, one study found molecular chaperones as mutant HTT interactors, in support of previous findings and highlighting the central role of these folding machineries as HTT modifiers [112].…”

Section: Proteomic Screeningmentioning

confidence: 74%

Experimental Models for Identifying Modifiers of Polyglutamine-Induced Aggregation and Neurodegeneration

Calamini

¹

,

Lo

²

,

Kaltenbach

³

2013

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Huntington's disease (HD) typifies a class of inherited neurodegenerative disorders in which a CAG expansion in a single gene leads to an extended polyglutamine tract and misfolding of the expressed protein, driving cumulative neural dysfunction and degeneration. HD is invariably fatal with symptoms that include progressive neuropsychiatric and cognitive impairments, and eventual motor disability. No curative therapies yet exist for HD and related polyglutamine diseases; therefore, substantial efforts have been made in the drug discovery field to identify potential drug and drug target candidates for disease-modifying treatment. In this context, we review here a range of early-stage screening approaches based in in vitro, cellular, and invertebrate models to identify pharmacological and genetic modifiers of polyglutamine aggregation and induced neurodegeneration. In addition, emerging technologies, including high-content analysis, threedimensional culture models, and induced pluripotent stem cells are increasingly being incorporated into drug discovery screening pipelines for protein misfolding disorders. Together, these diverse screening strategies are generating novel and exciting new probes for understanding the disease process and for furthering development of therapeutic candidates for eventual testing in the clinical setting.

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“…Recently, both WT and mHtt have been found to co-purify with several translation-related proteins and co-fractionate with ribosomes (Culver et al, 2012). Furthermore, the overexpression of Htt inhibits cap-dependent translation of a reporter mRNA in an in vitro system (Culver et al, 2012). During spermiogenesis, translational control is essential because de novo protein production is needed for the terminal steps of germ cell differentiation, which occur after transcription has ended.…”

Section: Discussionmentioning

confidence: 99%

Germline deletion of huntingtin causes male infertility and arrested spermiogenesis in mice

Yan¹,

Zhang²,

Liu³

et al. 2016

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Human Huntingtin (HTT), a Huntington's disease gene, is highly expressed in the mammalian brain and testis. Simultaneous knockout of mouse Huntingtin (Htt) in brain and testis impairs male fertility, providing evidence for a link between Htt and spermatogenesis; however, the underlying mechanism remains unclear. To understand better the function of Htt in spermatogenesis, we restricted the genetic deletion specifically to the germ cells using the Cre/loxP sitespecific recombination strategy and found that the resulting mice manifested smaller testes, azoospermia and complete male infertility. Meiotic chromosome spread experiments showed that the process of meiosis was normal in the absence of Htt. Notably, we found that Httdeficient round spermatids did not progress beyond step 3 during the post-meiotic phase, when round spermatids differentiate into mature spermatozoa. Using an iTRAQ-based quantitative proteomic assay, we found that knockout of Htt significantly altered the testis protein profile. The differentially expressed proteins exhibited a remarkable enrichment for proteins involved in translation regulation and DNA packaging, suggesting that Htt might play a role in spermatogenesis by regulating translation and DNA packaging in the testis.

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“…Eukaryotic translation initiation factor 2 (Eif2) signaling was predicted to be activated based on the upregulation of 17 ribosomal proteins. This translation activation is likely induced by knockout of Htt, as WT Htt functions with Ago2, the catalytic component of the RNAinduced-silencing complex, in a post-transcriptional gene silencing pathway, and its overexpression strongly inhibited translation in HeLa cell extracts (Savas et al, 2008;Culver et al, 2012).…”

Section: Functional Annotation Of Differentially Expressed Proteins Imentioning

confidence: 99%

“…Recently, both WT and mHtt have been found to co-purify with several translation-related proteins, and co-fractionate with ribosomes. Furthermore, Htt overexpression inhibits capdependent translation of a reporter mRNA in an in vitro system (Culver et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Germline deletion of huntingtin causes male infertility and arrested spermiogenesis in mice

Yan

¹

,

Zhang

²

,

Liu

³

et al. 2016

Journal of Cell Science

View full text Add to dashboard Cite

Human Huntingtin (HTT), a Huntington's disease gene, is highly expressed in the mammalian brain and testis. Simultaneous knockout of mouse Huntingtin (Htt) in brain and testis impairs male fertility, providing evidence for a link between Htt and spermatogenesis; however, the underlying mechanism remains unclear. To understand better the function of Htt in spermatogenesis, we restricted the genetic deletion specifically to the germ cells using the Cre/loxP sitespecific recombination strategy and found that the resulting mice manifested smaller testes, azoospermia and complete male infertility. Meiotic chromosome spread experiments showed that the process of meiosis was normal in the absence of Htt. Notably, we found that Httdeficient round spermatids did not progress beyond step 3 during the post-meiotic phase, when round spermatids differentiate into mature spermatozoa. Using an iTRAQ-based quantitative proteomic assay, we found that knockout of Htt significantly altered the testis protein profile. The differentially expressed proteins exhibited a remarkable enrichment for proteins involved in translation regulation and DNA packaging, suggesting that Htt might play a role in spermatogenesis by regulating translation and DNA packaging in the testis.

show abstract

Proteomic Analysis of Wild-type and Mutant Huntingtin-associated Proteins in Mouse Brains Identifies Unique Interactions and Involvement in Protein Synthesis

Cited by 124 publications

References 72 publications

Experimental Models for Identifying Modifiers of Polyglutamine-Induced Aggregation and Neurodegeneration

Experimental Models for Identifying Modifiers of Polyglutamine-Induced Aggregation and Neurodegeneration

Germline deletion of huntingtin causes male infertility and arrested spermiogenesis in mice

Germline deletion of huntingtin causes male infertility and arrested spermiogenesis in mice

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