Does N-terminal huntingtin function as a 'holdase' for inhibiting cellular protein aggregation?

Abstract

Proteolytic cleavage of huntingtin gives rise to N-terminal fragments. While the role of truncated mutant huntingtin is described in Huntington's disease (HD) pathogenesis, the function of N-terminal wild-type protein is less studied. The yeast model of HD is generated by the presence of FLAG tag and absence of polyproline tract as flanking sequences of the elongated polyglutamine stretch. We show that the same sequence derived from wild-type huntingtin exon1 is able to inhibit the aggregation of proteins in vitro and in yeast cells. It is able to stabilize client proteins as varied as luciferase, α-synuclein, and p53 in a soluble but non-native state. This is somewhat similar to the 'holdase' function of small heat shock proteins and 'nonchaperone proteins' which are able to stabilize partially unfolded client proteins in a nonspecific manner, slowing down their aggregation. Mutagenesis studies show this property to be localized at the N17 domain preceding the polyglutamine tract. Distortion of this ordered segment, either by deletion of this segment or mutation of a single residue (L4A), leads to decreased stability and increased aggregation of client proteins. It is interesting to note that the helical conformation of the N17 domain is also essential for aggregation of the N-terminal mutant protein. Our results provide evidence for a novel function for the amphipathic helix derived from exon1 of wild-type huntingtin.

• Authors: Ratnika Sethi, Neha Tripathi, Anusha R Pallapati, Abhishek Gaikar, Prasad V Bharatam, Ipsita Roy
• Journal: The FEBS journal (FEBS J) Vol. 285 Issue 10 Pg. 1791-1811 (05 2018) ISSN: 1742-4658 [Electronic] England
• PMID: 29630769 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 Federation of European Biochemical Societies.

Chemical References

• HTT protein, human
• Huntingtin Protein
• Peptides
• enhanced green fluorescent protein
• Green Fluorescent Proteins
• polyproline

Topics

• Amino Acid Sequence
• Exons
• Green Fluorescent Proteins (genetics)
• Humans
• Huntingtin Protein (chemistry, genetics, physiology)
• Hydrophobic and Hydrophilic Interactions
• Models, Biological
• Mutagenesis
• Particle Size
• Peptides (metabolism)
• Protein Aggregation, Pathological
• Protein Conformation
• Protein Stability
• Proteolysis
• Sequence Homology, Amino Acid
• Solubility