A role of cellular translation regulation associated with toxic Huntingtin protein


Huntington’s disease (HD) is a severe neurodegenerative disorder caused by poly Q repeat expansion in the Huntingtin (Htt) gene. While the Htt amyloid aggregates are known to affect many cellular processes, their role in translation has not been addressed. Here we report that pathogenic Htt expression causes a protein synthesis deficit in cells. We find a functional prion-like protein, the translation regulator Orb2, to be sequestered by Htt aggregates in cells. Co-expression of Orb2 can partially rescue the lethality associated with poly Q expanded Htt. These findings can be relevant for HD as human homologs of Orb2 are also sequestered by pathogenic Htt aggregates. Our work suggests that translation dysfunction is one of the contributors to the pathogenesis of HD and new therapies targeting protein synthesis pathways might help to alleviate disease symptoms.

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We acknowledge Prof. Troy Littleton, Prof. Kausik Si for sharing several Drosophila stocks and plasmids with us. We also thank Addgene and Bloomington Drosophila stock center for some of the plasmids and fly lines used in this work. We thank Prof. Kausik Si, Dr. Gunther Hollopeter, Dr. Irina Dudanova and Dr. Deepa Subramanyam for their suggestions on the work. We thank Dr. Vidisha Tripathi for letting us use the qPCR machine. A.M acknowledges Dr. Arvind Sahu, Dr. Vasudevan Seshadri and Dr. Ajay Pillai for their support. This work was supported by funding from Wellcome Trust-DBT India Alliance intermediate fellowship (IA/I/13/2/501030) to AM along with intramural funding from NCCS. TB was supported by Ramalingaswami fellowship from DBT (BT/RLF/Re-entry/54/2013) and IYBA Grant (BT/09/IYBA/2015/03).

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HJ started this project and identified Orb2 isoforms as rescuers of Htt pathogenicity. VG performed the S2 cell polysome and puromycin incorporation experiments. MS1 performed S2 cell imaging and FRAP experiments. MS2 did the Yeast polysome experiments. AR did the Orb2 rescue and Orb2 RNAi experiments. MD performed the Orb2 level quantitation, OPP staining, Htt clonings, and immunoprecipitation experiments. RC and AD performed the Yeast lethality and growth curve experiments. TB and AM conceived and designed the experiments. AM wrote the manuscript.

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Correspondence to Tania Bose or Amitabha Majumdar.

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FACS analysis of S2 cells transfected with HttQ15 and HttQ138 stained with 7AAD to check cell viability. No significant difference was observed with respect to 7AAD negative and RFP positive cells between HttQ15 and HttQ138 samples. Data is from 3 independent experiments and is represented as % live cells. Error bars represent SEM and significance is tested using unpaired one-tailed t-test (B) Cell counting of Trypan blue stained Yeast cells at 8 hours post galactose induction show no significant difference between cells expressing HttQ25 and HttQ103 (C) Yeast growth curve of HttQ25 and HttQ103 done with 3 independent repeats. At the 8 hour timepoint when polysome experiments were done the difference in growth in Q25 and Q103 is not significant (unpaired one-tailed t-test) with a p-value of 0.1363 (D) Quantitative real-time PCR to detect mRNA levels in Drosophila brains expressing HttQ15 and HttQ138 show no significant difference in endogenous transcript levels of Orb2. Data is from n=4 independent experiments and is represented as relative fold change in Orb2 levels for HttQ138 compared to HttQ15. Error bars represent SEM and significance is tested using unpaired one-tailed t-test and the p-value is 0.1661(ns) (E) Images of Neuro2A cells expressing RFP tagged HttQ15 and HttQ138. HttQ15 shows a diffused expression pattern while HttQ138 forms aggregate in these cell lines. 1 (PDF 2251 kb)

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Joag, H., Ghatpande, V., Desai, M. et al. A role of cellular translation regulation associated with toxic Huntingtin protein. Cell. Mol. Life Sci. 77, 3657–3670 (2020). https://doi.org/10.1007/s00018-019-03392-y

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  • Huntington’s disease
  • Functional-prion-like protein
  • Translation regulator
  • Orb2