Pharmaceutical Research

, Volume 29, Issue 4, pp 1098–1109

Changing the Subcellular Location of the Oncoprotein Bcr-Abl Using Rationally Designed Capture Motifs

  • Andrew S. Dixon
  • Jonathan E. Constance
  • Tomoyuki Tanaka
  • Terence H. Rabbitts
  • Carol S. Lim
Research Paper

DOI: 10.1007/s11095-011-0654-8

Cite this article as:
Dixon, A.S., Constance, J.E., Tanaka, T. et al. Pharm Res (2012) 29: 1098. doi:10.1007/s11095-011-0654-8

ABSTRACT

Purpose

Bcr-Abl, the causative agent of chronic myelogenous leukemia (CML), localizes in the cytoplasm where its oncogenic signaling leads to proliferation of cells. If forced into the nucleus Bcr-Abl causes apoptosis. To achieve nuclear translocation, binding domains for capture of Bcr-Abl were generated and attached to proteins with signals destined for the nucleus. These resulting proteins would be capable of binding and translocating endogenous Bcr-Abl to the nucleus.

Methods

Bcr-Abl was targeted at 3 distinct domains for capture: by construction of high affinity intracellular antibody domains (iDabs) to regions of Bcr-Abl known to promote cytoplasmic retention, via its coiled coil domain (CC), and through a naturally occurring protein-protein interaction domain (RIN1). These binding domains were then tested for their ability to escort Bcr-Abl into the nucleus using a “protein switch” or attachment of 4 nuclear localization signals (NLSs).

Results

Although RIN1, ABI7-iDab, and CCmut3 constructs all produced similar colocalization with Bcr-Abl, only 4NLS-CCmut3 produced efficient nuclear translocation of Bcr-Abl.

Conclusions

We demonstrate that a small binding domain can be used to control the subcellular localization of Bcr-Abl, which may have implications for CML therapy. Our ultimate future goal is to change the location of critical proteins to alter their function.

KEY WORDS

Bcr-Abl chronic myelogenous leukemia intracellular domain antibody subcellular targeting oncoprotein 

ABBREVIATIONS

Bcr-Abl

breakpoint cluster region/Abelson oncogene

CC

coiled coil

CML

chronic myelogenous leukemia

iDab

intracellular domain antibody

NLS

nuclear localization signal

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Andrew S. Dixon
    • 1
  • Jonathan E. Constance
    • 2
  • Tomoyuki Tanaka
    • 3
  • Terence H. Rabbitts
    • 3
  • Carol S. Lim
    • 1
  1. 1.Department of Pharmaceutics and Pharmaceutical Chemistry College of PharmacyUniversity of UtahSalt Lake CityUSA
  2. 2.Department of Pharmacology and Toxicology, College of PharmacyUniversity of UtahSalt Lake CityUSA
  3. 3.Section of Experimental Therapeutics Leeds Institute of Molecular MedicineSt. James’s University HospitalLeedsUK

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