A bidirectional system for the dynamic small molecule control of intracellular fusion proteins

Taavi K. Neklesa, Devin J. Noblin, Alexander Kuzin, Scott Lew, Jayaraman Seetharaman, Thomas B. Acton, Gregory Kornhaber, Rong Xiao, Gaetano T. Montelione, Liang Tong, Craig M. Crews

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Small molecule control of intracellular protein levels allows temporal and dose-dependent regulation of protein function. Recently, we developed a method to degrade proteins fused to a mutant dehalogenase (HaloTag2) using small molecule hydrophobic tags (HyTs). Here, we introduce a complementary method to stabilize the same HaloTag2 fusion proteins, resulting in a unified system allowing bidirectional control of cellular protein levels in a temporal and dose-dependent manner. From a small molecule screen, we identified N-(3,5-dichloro-2-ethoxybenzyl)-2H-tetrazol-5-amine as a nanomolar HALoTag2 Stabilizer (HALTS1) that reduces the Hsp70:HaloTag2 interaction, thereby preventing HaloTag2 ubiquitination. Finally, we demonstrate the utility of the HyT/HALTS system in probing the physiological role of therapeutic targets by modulating HaloTag2-fused oncogenic H-Ras, which resulted in either the cessation (HyT) or acceleration (HALTS) of cellular transformation. In sum, we present a general platform to study protein function, whereby any protein of interest fused to HaloTag2 can be either degraded 10-fold or stabilized 5-fold using two corresponding compounds.

Original languageEnglish (US)
Pages (from-to)2293-2300
Number of pages8
JournalACS chemical biology
Volume8
Issue number10
DOIs
StatePublished - Oct 18 2013

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine

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